8,974 results on '"Nucleotidyltransferases"'
Search Results
202. Pioneering technique reveals new layer of human gene regulation.
- Abstract
A new study led by researchers from NYU Grossman School of Medicine has developed a technique called Long Range Cleavage sequencing (LORAX-seq) that can detect and determine the frequency of a molecular event called "backtracking" in the genetic material of any species. Backtracking is a form of gene regulation that influences thousands of human genes, including those involved in cell division and development. The study found that backtracking occurs frequently throughout genomes, lasts longer than previously thought, and has functions beyond DNA repair. The researchers suggest that backtracking may be a widespread form of genetic regulation in various species. [Extracted from the article]
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- 2024
203. Karolinska Institute Researcher Details Findings in Nucleoproteins (Chromatin damage generated by DNA intercalators leads to degradation of RNA Polymerase II).
- Abstract
A recent study conducted at the Karolinska Institute in Stockholm, Sweden, has found that certain DNA intercalators, commonly used in cancer therapy, can impact chromatin biology by interfering with the stability of RNA polymerases I, II, and III. These intercalators induce degradation of RNA polymerase II and enable the trapping of topoisomerases on the chromatin. The study suggests that these compounds have a cumulative impact on general transcription machinery and may disrupt chromatin homeostasis, contributing to their cytotoxic effects. The research provides valuable insights into the mechanisms of DNA intercalators and their potential implications for cancer treatment. [Extracted from the article]
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- 2024
204. Data on Genomics and Genetics Discussed by Researchers at Columbia University (Bacterial Genome Engineering Using Crispr-associated Transposases).
- Abstract
Researchers at Columbia University have discussed the use of CRISPR-associated transposases for bacterial genome engineering. These transposons have the potential to revolutionize genome engineering by integrating large genetic payloads with high accuracy and programmability, without the need for homologous recombination machinery. The researchers have developed a detailed protocol for engineering bacterial genomes using CRISPR-associated transposase (CAST) systems, including guidelines for vector selection, customization of guide RNAs and DNA payloads, delivery methods, and genotypic analysis. This method allows for the isolation of clonal strains with specific genomic integration events within 1-2 weeks. The research has been peer-reviewed and published in Nature Protocols. [Extracted from the article]
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- 2024
205. Study Findings from Yangzhou University Provide New Insights into Circovirus (DEAD-box RNA helicase 21 interacts with porcine circovirus type 2 Cap protein and facilitates viral replication).
- Abstract
A study conducted by researchers at Yangzhou University in China has found that the interaction between DEAD-box RNA helicase 21 (DDX21) and the capsid (Cap) protein of porcine circovirus type 2 (PCV2) plays a crucial role in the replication of the virus. The researchers discovered that PCV2 infection causes the relocation of DDX21 from the nucleolus to the cytoplasm in cultured PK-15 cells. They also found that the nuclear localization signal (NLS) of PCV2 Cap directly interacts with DDX21. Depletion of DDX21 resulted in impaired PCV2 replication, while overexpression of DDX21 increased PCV2 replication. These findings provide insights into potential targets for the prevention and control of PCV2 infection. [Extracted from the article]
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- 2024
206. Patent Issued for Contiguity preserving transposition (USPTO 11873480).
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- 2024
207. The RNA polymerase II general transcription factor TFIIB is a target for transcriptome control during cellular stress and viral infection.
- Abstract
A recent preprint abstract discusses the mechanisms underlying transcriptional repression during cellular stress and viral infection. The study focuses on the RNA polymerase II general transcription factor TFIIB, which is targeted for post-translational turnover by two pathways during stress. The researchers found that TFIIB is cleaved by activated caspase-3 and targeted for rapid proteasome-mediated turnover by the E3 ubiquitin ligase TRIM28. The study suggests that TFIIB may play a crucial role in transcriptional outcomes during stress and could be a key target for nuclear replicating DNA viruses. However, it is important to note that this preprint has not been peer-reviewed. [Extracted from the article]
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- 2024
208. ANGEL2 is a member of the CCR4 family of deadenylases with 2′,3′-cyclic phosphatase activity.
- Author
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Pinto, Paola H., Kroupova, Alena, Schleiffer, Alexander, Mechtler, Karl, Jinek, Martin, Weitzer, Stefan, and Martinez, Javier
- Subjects
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TRANSFER RNA , *MESSENGER RNA , *NUCLEOTIDYLTRANSFERASES , *RIBOSOMES , *ENDONUCLEASES - Abstract
RNA molecules are frequently modified with a terminal 2′,3′-cyclic phosphate group as a result of endonuclease cleavage, exonuclease trimming, or de novo synthesis. During pre-transfer RNA (tRNA) and unconventional messenger RNA (mRNA) splicing, 2′,3′-cyclic phosphates are substrates of the tRNA ligase complex, and their removal is critical for recycling of tRNAs upon ribosome stalling. We identified the predicted deadenylase angel homolog 2 (ANGEL2) as a human phosphatase that converts 2′,3′-cyclic phosphates into 2′,3′-OH nucleotides. We analyzed ANGEL2’s substrate preference, structure, and reaction mechanism. Perturbing ANGEL2 expression affected the efficiency of pre-tRNA processing, X-box–binding protein 1 (XBP1) mRNA splicing during the unfolded protein response, and tRNA nucleotidyltransferase 1 (TRNT1)–mediated CCA addition onto tRNAs. Our results indicate that ANGEL2 is involved in RNA pathways that rely on the ligation or hydrolysis of 2′,3′-cyclic phosphates. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
209. Anti‐MDA5‐associated dermatomyositis.
- Author
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Gupta, Rajiv, Kumar, Sunil, Gow, Peter, (Hsien‐Cheng) Chang, Leon, and Yen, Linda
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THERAPEUTIC use of immunoglobulins , *AUTOANTIBODIES , *CYCLOSPORINE , *DERMATOMYOSITIS , *IMMUNOSUPPRESSIVE agents , *INTERSTITIAL lung diseases , *TREATMENT effectiveness , *CYCLOPHOSPHAMIDE , *METHYLPREDNISOLONE , *NUCLEOTIDYLTRANSFERASES - Abstract
Anti‐MDA5‐associated dermatomyositis (MDA5‐associated DM) is an uncommon presentation of idiopathic inflammatory myositis, typically amyopathic, associated with rapidly progressive, treatment refractory interstitial lung disease and poor prognosis, particularly in patients with concomitant rapidly progressive interstitial lung disease (RP‐ILD). We report two cases of MDA5‐associated DM with fatal outcome in one of the patients, despite 'aggressive triple therapy' for RP‐ILD. [ABSTRACT FROM AUTHOR]
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- 2020
- Full Text
- View/download PDF
210. Determinants of Replication-Fork Pausing at tRNA Genes in Saccharomyces cerevisiae.
- Author
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Yeung, Rani and Smith, Duncan J.
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DNA , *GENOMES , *PROTEINS , *SACCHAROMYCES , *TRANSCRIPTION factors , *TRANSFER RNA , *YEAST , *DNA-binding proteins , *NUCLEOTIDYLTRANSFERASES , *CHROMOSOME structure - Abstract
Transfer RNA (tRNA) genes are widely studied sites of replication-fork pausing and genome instability in the budding yeast Saccharomyces cerevisiae. tRNAs are extremely highly transcribed and serve as constitutive condensin binding sites. tRNA transcription by RNA polymerase III has previously been identified as stimulating replication-fork pausing at tRNA genes, but the nature of the block to replication has not been incontrovertibly demonstrated. Here, we describe a systematic, genome-wide analysis of the contributions of candidates to replication-fork progression at tDNAs in yeast: transcription factor binding, transcription, topoisomerase activity, condensin-mediated clustering, and Rad18-dependent DNA repair. We show that an asymmetric block to replication is maintained even when tRNA transcription is abolished by depletion of one or more subunits of RNA polymerase III. By contrast, analogous depletion of the essential transcription factor TFIIIB removes the obstacle to replication. Therefore, our data suggest that the RNA polymerase III transcription complex itself represents an asymmetric obstacle to replication even in the absence of RNA synthesis. We additionally demonstrate that replication-fork progression past tRNA genes is unaffected by the global depletion of condensin from the nucleus, and can be stimulated by the removal of topoisomerases or Rad18-dependent DNA repair pathways. [ABSTRACT FROM AUTHOR]
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- 2020
- Full Text
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211. Optimal Growth Temperature and Intergenic Distances in Bacteria, Archaea, and Plastids of Rhodophytic Branch.
- Author
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Lyubetsky, Vassily A., Zverkov, Oleg A., Rubanov, Lev I., and Seliverstov, Alexandr V.
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RNA analysis , *ALGAE , *BACTERIA , *CYTOPLASM , *GENOMES , *MICROBIAL ecology , *REGRESSION analysis , *TEMPERATURE , *NUCLEOTIDYLTRANSFERASES - Abstract
The lengths of intergenic regions between neighboring genes that are convergent, divergent, or unidirectional were calculated for plastids of the rhodophytic branch and complete archaeal and bacterial genomes. Statistically significant linear relationships between any pair of the medians of these three length types have been revealed in each genomic group. Exponential relationships between the optimal growth temperature and each of the three medians have been revealed as well. The leading coefficients of the regression equations relating all pairs of the medians as well as temperature and any of the medians have the same sign and order of magnitude. The results obtained for plastids, archaea, and bacteria are also similar at the qualitative level. For instance, the medians are always low at high temperatures. At low temperatures, the medians tend to statistically significant greater values and scattering. The original model was used to test our hypothesis that the intergenic distances are optimized in particular to decrease the competition of RNA polymerases within the locus that results in transcribing shortened RNAs. Overall, this points to an effect of temperature for both remote and close genomes. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
212. Inhibitory complex of the transmembrane ammonia channel, AmtB, and the cytosolic regulatory protein, GlnK, at 1.96 Å
- Author
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Gruswitz, Franz, O'Connell, Joseph, and Stroud, Robert M
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Emerging Infectious Diseases ,Adenosine Triphosphate ,Cation Transport Proteins ,Escherichia coli Proteins ,Ketoglutaric Acids ,Nucleotidyltransferases ,PII Nitrogen Regulatory Proteins ,Periplasm ,Protein Conformation ,Protein Processing ,Post-Translational ,Quaternary Ammonium Compounds ,membrane ,regulation ,structure ,allosteric ,dinucleotide - Abstract
Ammonia conductance is highly regulated. A P(II) signal transduction protein, GlnK, is the final regulator of transmembrane ammonia conductance by the ammonia channel AmtB in Escherichia coli. The complex formed between AmtB and inhibitory GlnK at 1.96-A resolution shows that the trimeric channel is blocked directly by GlnK and how, in response to intracellular nitrogen status, the ability of GlnK to block the channel is regulated by uridylylation/deuridylylation at Y51. ATP and Mg(2+) augment the interaction of GlnK. The hydrolyzed product, adenosine 5'-diphosphate orients the surface of GlnK for AmtB blockade. 2-Oxoglutarate diminishes AmtB/GlnK association, and sites for 2-oxoglutarate are evaluated.
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- 2007
213. Design, Synthesis, and Biochemical and Biological Evaluation of Novel 7-Deazapurine Cyclic Dinucleotide Analogues as STING Receptor Agonists
- Author
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Zdeněk Vavřina, Pavla Perlíková, Nemanja Milisavljević, Florian Chevrier, Miroslav Smola, Joshua Smith, Milan Dejmek, Vojtěch Havlíček, Miloš Buděšínský, Radek Liboska, Lenka Vaneková, Jiří Brynda, Evzen Boura, Pavlína Řezáčová, Michal Hocek, and Gabriel Birkuš
- Subjects
Mice ,Drug Discovery ,Animals ,Humans ,Membrane Proteins ,Cytokines ,Molecular Medicine ,Interferons ,Nucleotides, Cyclic ,Ligands ,Nucleotidyltransferases - Abstract
Cyclic dinucleotides (CDNs) are second messengers that activate stimulator of interferon genes (STING). The cGAS-STING pathway plays a promising role in cancer immunotherapy. Here, we describe the synthesis of CDNs containing 7-substituted 7-deazapurine moiety. We used mouse cyclic GMP-AMP synthase and bacterial dinucleotide synthases for the enzymatic synthesis of CDNs. Alternatively, 7-(het)aryl 7-deazapurine CDNs were prepared by Suzuki-Miyaura cross-couplings. New CDNs were tested in biochemical and cell-based assays for their affinity to human STING. Eight CDNs showed better activity than 2'3'
- Published
- 2022
214. Crystallographic analysis of engineered polymerases synthesizing phosphonomethylthreosyl nucleic acid
- Author
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Mohammad Hajjar, Nicholas Chim, Chao Liu, Piet Herdewijn, and John C Chaput
- Subjects
Biochemistry & Molecular Biology ,Science & Technology ,NUCLEOSIDES ,Nucleotides ,Polymers ,GENETIC POLYMERS ,DNA ,Nucleotidyltransferases ,Nucleic Acids ,REPLICATION ,Genetics ,Nucleic Acid Conformation ,TOOL ,RNA ,TNA ,Life Sciences & Biomedicine ,DUPLEX ,SYSTEM ,FORM ,DNA Primers - Abstract
Xeno-nucleic acids (XNAs) are synthetic genetic polymers with backbone structures composed of non-ribose or non-deoxyribose sugars. Phosphonomethylthreosyl nucleic acid (pTNA), a type of XNA that does not base pair with DNA or RNA, has been suggested as a possible genetic material for storing synthetic biology information in cells. A critical step in this process is the synthesis of XNA episomes using laboratory-evolved polymerases to copy DNA information into XNA. Here, we investigate the polymerase recognition of pTNA nucleotides using X-ray crystallography to capture the post-catalytic complex of engineered polymerases following the sequential addition of two pTNA nucleotides onto the 3'-end of a DNA primer. High-resolution crystal structures reveal that the polymerase mediates Watson-Crick base pairing between the extended pTNA adducts and the DNA template. Comparative analysis studies demonstrate that the sugar conformation and backbone position of pTNA are structurally more similar to threose nucleic acid than DNA even though pTNA and DNA share the same six-atom backbone repeat length. Collectively, these findings provide new insight into the structural determinants that guide the enzymatic synthesis of an orthogonal genetic polymer, and may lead to the discovery of new variants that function with enhanced activity. ispartof: NUCLEIC ACIDS RESEARCH vol:50 issue:17 pages:9663-9674 ispartof: location:England status: published
- Published
- 2022
215. Human Thg1 displays tRNA-inducible GTPase activity
- Author
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Titi Rindi Antika, Kun Rohmatan Nazilah, Yi-Hsueh Lee, Ya-Ting Lo, Chung-Shu Yeh, Fu-Lung Yeh, Tien-Hsien Chang, Tzu-Ling Wang, and Chien-Chia Wang
- Subjects
Adenosine Triphosphate ,Guanosine ,RNA, Transfer ,Genetics ,Humans ,Guanosine Triphosphate ,Saccharomyces cerevisiae ,RNA, Transfer, His ,Nucleotidyltransferases ,GTP Phosphohydrolases ,Histidine-tRNA Ligase - Abstract
tRNAHis guanylyltransferase (Thg1) catalyzes the 3′-5′ incorporation of guanosine into position -1 (G-1) of tRNAHis. G-1 is unique to tRNAHis and is crucial for recognition by histidyl-tRNA synthetase (HisRS). Yeast Thg1 requires ATP for G-1 addition to tRNAHis opposite A73, whereas archaeal Thg1 requires either ATP or GTP for G-1 addition to tRNAHis opposite C73. Paradoxically, human Thg1 (HsThg1) can add G-1 to tRNAsHis with A73 (cytoplasmic) and C73 (mitochondrial). As N73 is immediately followed by a CCA end (positions 74–76), how HsThg1 prevents successive 3′-5′ incorporation of G-1/G-2/G-3 into mitochondrial tRNAHis (tRNAmHis) through a template-dependent mechanism remains a puzzle. We showed herein that mature native human tRNAmHis indeed contains only G-1. ATP was absolutely required for G-1 addition to tRNAmHis by HsThg1. Although HsThg1 could incorporate more than one GTP into tRNAmHisin vitro, a single-GTP incorporation prevailed when the relative GTP level was low. Surprisingly, HsThg1 possessed a tRNA-inducible GTPase activity, which could be inhibited by ATP. Similar activity was found in other high-eukaryotic dual-functional Thg1 enzymes, but not in yeast Thg1. This study suggests that HsThg1 may downregulate the level of GTP through its GTPase activity to prevent multiple-GTP incorporation into tRNAmHis.
- Published
- 2022
216. Inhibition of cGAS in Paraventricular Nucleus Attenuates Hypertensive Heart Injury Via Regulating Microglial Autophagy
- Author
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Chengzhi Han, Xinyi Qian, Xiaorong Ren, Shutian Zhang, Li Hu, Jingyao Li, Yijun Huang, Renhui Huang, Kokwin Ooi, Hong Lin, and Chunmei Xia
- Subjects
Sirolimus ,Heart Diseases ,Angiotensin II ,Neuroscience (miscellaneous) ,DNA, Mitochondrial ,Nucleotidyltransferases ,Mice ,Cellular and Molecular Neuroscience ,Heart Injuries ,Neurology ,Hypertension ,Autophagy ,Animals ,Microglia ,Paraventricular Hypothalamic Nucleus - Abstract
Neuroinflammation in the cardiovascular center plays a critical role in the progression of hypertensive heart disease. And microglial autophagy is involved in the regulation of neuroinflammation. Cyclic GMP-AMP synthase (cGAS), a cytosolic DNA sensor, senses mitochondrial DNA (mtDNA) and regulates autophagy. The detailed mechanisms of central cGAS affects neuroinflammatory response in hypertensive heart disease via regulating autophagy remain unknown. Angiotensin II (Ang II, 1.5 mg·kg
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- 2022
217. Microglial <scp>AIM2</scp> alleviates antiviral‐related neuro‐inflammation in mouse models of Parkinson's disease
- Author
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Wen‐Juan Rui, Sheng Li, Lin Yang, Ying Liu, Yi Fan, Ying‐Chao Hu, Chun‐Mei Ma, Bing‐Wei Wang, and Jing‐Ping Shi
- Subjects
Inflammation ,Pyrrolidines ,Inflammasomes ,Dopaminergic Neurons ,Mice, Transgenic ,Parkinson Disease ,Antiviral Agents ,Nucleotidyltransferases ,DNA-Binding Proteins ,Mice, Inbred C57BL ,Disease Models, Animal ,Mice ,Cellular and Molecular Neuroscience ,Neurology ,1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine ,Animals ,RNA ,Interferon Regulatory Factor-3 ,Microglia ,Melanoma ,Proto-Oncogene Proteins c-akt - Abstract
Inflammasome involvement in Parkinson's disease (PD) has been intensively investigated. Absent in melanoma 2 (AIM2) is an essential inflammasome protein known to contribute to the development of several neurological diseases. However, a specific role for AIM2 in PD has not been reported. In this study, we investigated the effect of AIM2 in the N-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP)-induced PD model by use of various knockout and bone marrow chimeric mice. The mechanism of action for AIM2 in PD was assessed by RNA-sequencing and in vitro primary microglial transfection. Results were validated in the A30P transgenic mouse model of PD. In the MPTP mouse model, AIM2 activation was found to negatively regulate neuro-inflammation independent of the inflammasome. Microglial AIM2 deficiency exacerbated behavioral and pathological features of both MPTP-induced and transgenic PD mouse models. Mechanistically, AIM2 reduced cyclic GMP-AMP synthase (cGAS)-mediated antiviral-related inflammation by inhibition of AKT-interferon regulatory factor 3 (IRF3) phosphorylation. These results demonstrate microglial AIM2 to inhibit the antiviral-related neuro-inflammation associated with PD and provide for a foundation upon which to identify new therapeutic targets for treatment of the disease.
- Published
- 2022
218. Biodegradable MnO-Based Nanoparticles with Engineering Surface for Tumor Therapy: Simultaneous Fenton-Like Ion Delivery and Immune Activation
- Author
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Zhaoli Sun, Zhiyi Wang, Tao Wang, Jingjing Wang, Hongtao Zhang, Ziyuan Li, Shuren Wang, Fugeng Sheng, Jing Yu, and Yanglong Hou
- Subjects
Manganese Compounds ,Neoplasms ,Cell Line, Tumor ,Programmed Cell Death 1 Receptor ,General Engineering ,Humans ,Nanoparticles ,General Physics and Astronomy ,General Materials Science ,Reactive Oxygen Species ,Nucleotidyltransferases - Abstract
Immune checkpoint inhibitors have achieved significant clinical success but are still suffering from inadequate immune activation. It is worth noting that manganese as a nutritional inorganic trace element is closely associated with immune activation to fight against tumor growth and metastasis
- Published
- 2022
219. New mechanism of nephrotoxicity of triptolide: Oxidative stress promotes cGAS-STING signaling pathway
- Author
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Jun, Lu, Yi, Zhang, Huiyue, Dong, Jingjing, Sun, Ling, Zhu, Pengyang, Liu, Fuli, Wen, and Rong, Lin
- Subjects
Mice ,Oxidative Stress ,Drug-Related Side Effects and Adverse Reactions ,Physiology (medical) ,Animals ,Epoxy Compounds ,Humans ,Membrane Proteins ,Diterpenes ,Phenanthrenes ,Nucleotidyltransferases ,Biochemistry ,Signal Transduction - Abstract
Triptolide (TPL) is a bioactive component extracted from the traditional Chinese herb Tripterygium wilfordii Hook F., and has multiple pharmacological activities, such as anti-tumor activity. However, severe adverse effects and toxicity, especially nephrotoxicity, limit its clinical application. It has been demonstrated that mitochondrial defect is a major toxic effects of TPL. In this study, we show that triptolide activated the cGAS-STING signaling pathway in kidney tubular cells in vivo and in vitro. Renal injury models were established in BALB/c mice and human tubular epithelial cells using TPL. We found that TPL enhanced the phosphorylation levels of STING, TBK1 and IRF3, and upregulated the expression of IFNβ, which is the production of cGAS-STING signaling pathway. STING inhibitor C176 had protective effects in TPL-induced nephrocyte damage. STING siRNA down regulated the expression level of IFNβ. In addition, triptolide induced an increase in protein levels of the transcription factor BACH1, while transcriptional expression of the antioxidant enzyme HMOX1 was reduced due to the increased expression of BACH1. Furthermore, oxidative stress-induced mtDNA damage and DNA leakage caused activation of the cGAS-STING signaling pathway. Altogether, cGAS-STING signaling pathway involved in TPL induced nephrotoxicity. Inhibiting cGAS-STING over-activation may be a new strategy for alleviating renal injury of triptolide.
- Published
- 2022
220. Duck Enteritis Virus Inhibits the cGAS-STING DNA-Sensing Pathway To Evade the Innate Immune Response
- Author
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Li Gao, Rui Liu, Fuchun Yang, Xiaohan Li, Changjun Liu, Xiaole Qi, Hongyu Cui, Yanping Zhang, Suyan Wang, Xiaomei Wang, Yulong Gao, and Kai Li
- Subjects
Interferon Regulatory Factor-7 ,Immunology ,Alphaherpesvirinae ,Microbiology ,Nucleotidyltransferases ,Enteritis ,Immunity, Innate ,Virus-Cell Interactions ,Viral Proteins ,Ducks ,Virology ,Insect Science ,DNA, Viral ,Animals ,Signal Transduction ,Immune Evasion - Abstract
Cyclic GMP-AMP synthase (cGAS), a key DNA sensor, detects cytosolic viral DNA and activates the adaptor protein stimulator of interferon genes (STING) to initiate interferon (IFN) production and host innate antiviral responses. Duck enteritis virus (DEV) is a duck alphaherpesvirus that causes an acute and contagious disease with high mortality in waterfowl. In the present study, we found that DEV inhibits host innate immune responses during the late phase of viral infection. Furthermore, we screened DEV proteins for their ability to inhibit the cGAS-STING DNA-sensing pathway and identified multiple viral proteins, including UL41, US3, UL28, UL53, and UL24, which block IFN-β activation through this pathway. The DEV tegument protein UL41, which exhibited the strongest inhibitory effect, selectively downregulated the expression of interferon regulatory factor 7 (IRF7) by reducing its mRNA accumulation, thereby inhibiting the DNA-sensing pathway. Ectopic expression of UL41 markedly reduced viral DNA-triggered IFN-β production and promoted viral replication, whereas deficiency of UL41 in the context of DEV infection increased the IFN-β response to DEV and suppressed viral replication. In addition, ectopic expression of IRF7 inhibited the replication of the UL41-deficient virus, whereas IRF7 knockdown facilitated its replication. This study is the first report identifying multiple viral proteins encoded by a duck DNA virus, which inhibit the cGAS-STING DNA-sensing pathway. These findings expand our knowledge of DNA sensing in ducks and reveal a mechanism through which DEV antagonizes the host innate immune response. IMPORTANCE Duck enteritis virus (DEV) is a duck alphaherpesvirus that causes an acute and contagious disease with high mortality, resulting in substantial economic losses in the commercial waterfowl industry. The evasion of DNA-sensing pathway-mediated antiviral innate immunity is essential for the persistent infection and replication of many DNA viruses. However, the mechanisms used by DEV to modulate the DNA-sensing pathway remain poorly understood. In the present study, we found that DEV encodes multiple viral proteins to inhibit the cGAS-STING DNA-sensing pathway. The DEV tegument protein UL41 selectively diminished the accumulation of interferon regulatory factor 7 (IRF7) mRNA, thereby inhibiting the DNA-sensing pathway. Loss of UL41 potently enhanced the IFN-β response to DEV and impaired viral replication in ducks. These findings provide insights into the host-virus interaction during DEV infection and help develop new live attenuated vaccines against DEV.
- Published
- 2023
221. Phospholipase D3 degrades mitochondrial DNA to regulate nucleotide signaling and APP metabolism
- Author
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Zoë P. Van Acker, Anika Perdok, Ruben Hellemans, Katherine North, Inge Vorsters, Cedric Cappel, Jonas Dehairs, Johannes V. Swinnen, Ragna Sannerud, Marine Bretou, Markus Damme, and Wim Annaert
- Subjects
Multidisciplinary ,Nucleotides ,Phospholipases ,General Physics and Astronomy ,Chromogranin A ,Amyloidogenic Proteins ,General Chemistry ,DNA, Mitochondrial ,Nucleotidyltransferases ,General Biochemistry, Genetics and Molecular Biology ,Mitochondria - Abstract
Phospholipase D3 (PLD3) polymorphisms are linked to late-onset Alzheimer's disease (LOAD). Being a lysosomal 5'-3' exonuclease, its neuronal substrates remained unknown as well as how a defective lysosomal nucleotide catabolism connects to AD-proteinopathy. We identified mitochondrial DNA (mtDNA) as a major physiological substrate and show its manifest build-up in lysosomes of PLD3-defective cells. mtDNA accretion creates a degradative (proteolytic) bottleneck that presents at the ultrastructural level as a marked abundance of multilamellar bodies, often containing mitochondrial remnants, which correlates with increased PINK1-dependent mitophagy. Lysosomal leakage of mtDNA to the cytosol activates cGAS-STING signaling that upregulates autophagy and induces amyloid precursor C-terminal fragment (APP-CTF) and cholesterol accumulation. STING inhibition largely normalizes APP-CTF levels, whereas an APP knockout in PLD3-deficient backgrounds lowers STING activation and normalizes cholesterol biosynthesis. Collectively, we demonstrate molecular cross-talks through feedforward loops between lysosomal nucleotide turnover, cGAS-STING and APP metabolism that, when dysregulated, result in neuronal endolysosomal demise as observed in LOAD. ispartof: NATURE COMMUNICATIONS vol:14 issue:1 ispartof: location:England status: published
- Published
- 2023
222. IP6-stabilised HIV capsids evade cGAS/STING-mediated host immune sensing
- Author
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Papa, Guido, Albecka, Anna, Mallery, Donna, Vaysburd, Marina, Renner, Nadine, James, Leo C, Papa, Guido [0000-0002-5215-0014], Albecka, Anna [0000-0002-3672-5498], Mallery, Donna [0000-0003-2713-5215], James, Leo C [0000-0003-2131-0334], and Apollo - University of Cambridge Repository
- Subjects
capsid stability ,IP6 ,Capsid ,Host-Pathogen Interactions ,HIV ,Humans ,Membrane Proteins ,HIV Infections ,innate immunity ,Nucleotidyltransferases ,Immunity, Innate ,DNA sensing - Abstract
HIV-1 uses inositol hexakisphosphate (IP6) to build a metastable capsid capable of delivering its genome into the host nucleus. Here, we show that viruses that are unable to package IP6 lack capsid protection and are detected by innate immunity, resulting in the activation of an antiviral state that inhibits infection. Disrupting IP6 enrichment results in defective capsids that trigger cytokine and chemokine responses during infection of both primary macrophages and T-cell lines. Restoring IP6 enrichment with a single mutation rescues the ability of HIV-1 to infect cells without being detected. Using a combination of capsid mutants and CRISPR-derived knockout cell lines for RNA and DNA sensors, we show that immune sensing is dependent upon the cGAS-STING axis and independent of capsid detection. Sensing requires the synthesis of viral DNA and is prevented by reverse transcriptase inhibitors or reverse transcriptase active-site mutation. These results demonstrate that IP6 is required to build capsids that can successfully transit the cell and avoid host innate immune sensing.
- Published
- 2023
223. All four Mycobacterium tuberculosis glnA genes encode glutamine synthetase activities but only GlnA1 is abundantly expressed and essential for bacterial homeostasis
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Harth, Günter, Masleša‐Galić, Saša, Tullius, Michael V, and Horwitz, Marcus A
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Biochemistry and Cell Biology ,Biological Sciences ,Orphan Drug ,Prevention ,Infectious Diseases ,Rare Diseases ,Tuberculosis ,HIV/AIDS ,Biotechnology ,Genetics ,Infection ,Good Health and Well Being ,Amino Acid Isomerases ,Enzyme Inhibitors ,Gene Deletion ,Gene Expression Regulation ,Bacterial ,Gene Order ,Genes ,Bacterial ,Genes ,Essential ,Glutamate-Ammonia Ligase ,Glutamate-Cysteine Ligase ,Glutamine ,Methionine Sulfoximine ,Multigene Family ,Mutagenesis ,Insertional ,Mycobacterium smegmatis ,Mycobacterium tuberculosis ,Nucleotidyltransferases ,Peptide Synthases ,RNA ,Bacterial ,RNA ,Messenger ,Transcription Initiation Site ,Transcription ,Genetic ,Agricultural and Veterinary Sciences ,Medical and Health Sciences ,Microbiology ,Biological sciences - Abstract
Glutamine synthetases (GS) are ubiquitous enzymes that play a central role in every cell's nitrogen metabolism. We have investigated the expression and activity of all four genomic Mycobacterium tuberculosis GS - GlnA1, GlnA2, GlnA3 and GlnA4 - and four enzymes regulating GS activity and/or nitrogen and glutamate metabolism - adenylyl transferase (GlnE), gamma-glutamylcysteine synthase (GshA), UDP-N-acetylmuramoylalanine-D-glutamate ligase (MurD) and glutamate racemase (MurI). All eight genes are located in multigene operons except for glnA1, and all are transcribed in M. tuberculosis; however, some are not translated or translated at such low levels that the enzymes escape detection. Of the four GS, only GlnA1 can be detected. Each of the eight genes, as well as the glnA1-glnE-glnA2 cluster, was expressed separately in Mycobacterium smegmatis, and its gene product was characterized and assayed for enzymatic activity by analysing the reaction products. In M. smegmatis, all four recombinant-overexpressed GS are multimeric enzymes exhibiting GS activity. Whereas GlnA1, GlnA3 and GlnA4 catalyse the synthesis of L-glutamine, GlnA2 catalyses the synthesis of D-glutamine and D-isoglutamine. The generation of mutants in M. tuberculosis of the four glnA genes, murD and murI demonstrated that all of these genes except glnA1 are nonessential for in vitro growth. L-methionine-S,R-sulphoximine (MSO), previously demonstrated to inhibit M. tuberculosis growth in vitro and in vivo, strongly inhibited all four GS enzymes; hence, the design of MSO analogues with an improved therapeutic to toxic ratio remains a promising strategy for the development of novel anti-M. tuberculosis drugs.
- Published
- 2005
224. Characterization of the Genes Encoding the Cytosolic and Plastidial Forms of ADP-Glucose Pyrophosphorylase in Wheat Endosperm
- Author
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Burton, Rachel A, Johnson, Philip E, Beckles, Diane M, Fincher, Geoffrey B, Jenner, Helen L, Naldrett, Mike J, and Denyer, Kay
- Subjects
Biochemistry and Cell Biology ,Biological Sciences ,Genetics ,Amino Acid Sequence ,Base Sequence ,Chromatography ,Ion Exchange ,Cloning ,Molecular ,Cytosol ,DNA ,Complementary ,DNA ,Plant ,Gene Expression Regulation ,Developmental ,Gene Expression Regulation ,Enzymologic ,Gene Expression Regulation ,Plant ,Glucose-1-Phosphate Adenylyltransferase ,Molecular Sequence Data ,Nucleotidyltransferases ,Phylogeny ,Plastids ,Seeds ,Sequence Analysis ,DNA ,Sequence Analysis ,Protein ,Sequence Homology ,Amino Acid ,Sequence Homology ,Nucleic Acid ,Triticum ,Agricultural and Veterinary Sciences ,Plant Biology & Botany ,Plant biology - Abstract
In most species, the synthesis of ADP-glucose (Glc) by the enzyme ADP-Glc pyrophosphorylase (AGPase) occurs entirely within the plastids in all tissues so far examined. However, in the endosperm of many, if not all grasses, a second form of AGPase synthesizes ADP-Glc outside the plastid, presumably in the cytosol. In this paper, we show that in the endosperm of wheat (Triticum aestivum), the cytosolic form accounts for most of the AGPase activity. Using a combination of molecular and biochemical approaches to identify the cytosolic and plastidial protein components of wheat endosperm AGPase we show that the large and small subunits of the cytosolic enzyme are encoded by genes previously thought to encode plastidial subunits, and that a gene, Ta.AGP.S.1, which encodes the small subunit of the cytosolic form of AGPase, also gives rise to a second transcript by the use of an alternate first exon. This second transcript encodes an AGPase small subunit with a transit peptide. However, we could not find a plastidial small subunit protein corresponding to this transcript. The protein sequence of the purified plastidial small subunit does not match precisely to that encoded by Ta.AGP.S.1 or to the predicted sequences of any other known gene from wheat or barley (Hordeum vulgare). Instead, the protein sequence is most similar to those of the plastidial small subunits from chickpea (Cicer arietinum) and maize (Zea mays) and rice (Oryza sativa) seeds. These data suggest that the gene encoding the major plastidial small subunit of AGPase in wheat endosperm has yet to be identified.
- Published
- 2002
225. Functional screening reveals HORMAD1-driven gene dependencies associated with translesion synthesis and replication stress tolerance
- Author
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Dalia Tarantino, Callum Walker, Daniel Weekes, Helen Pemberton, Kathryn Davidson, Gonzalo Torga, Jessica Frankum, Ana M. Mendes-Pereira, Cynthia Prince, Riccardo Ferro, Rachel Brough, Stephen J. Pettitt, Christopher J. Lord, Anita Grigoriadis, and Andrew NJ Tutt
- Subjects
DNA Replication ,Cancer Research ,DNA Repair ,Phosphoric Diester Hydrolases ,Cell Cycle Proteins ,Triple Negative Breast Neoplasms ,DNA-Directed DNA Polymerase ,Nucleotidyltransferases ,Genomic Instability ,DNA-Binding Proteins ,X-ray Repair Cross Complementing Protein 1 ,Genetics ,Humans ,Molecular Biology ,DNA Damage - Abstract
HORMAD1 expression is usually restricted to germline cells, but it becomes mis-expressed in epithelial cells in ~60% of triple-negative breast cancers (TNBCs), where it is associated with elevated genomic instability (1). HORMAD1 expression in TNBC is bimodal with HORMAD1-positive TNBC representing a biologically distinct disease group. Identification of HORMAD1-driven genetic dependencies may uncover novel therapies for this disease group. To study HORMAD1-driven genetic dependencies, we generated a SUM159 cell line model with doxycycline-inducible HORMAD1 that replicated genomic instability phenotypes seen in HORMAD1-positive TNBC (1). Using small interfering RNA screens, we identified candidate genes whose depletion selectively inhibited the cellular growth of HORMAD1-expressing cells. We validated five genes (ATR, BRIP1, POLH, TDP1 and XRCC1), depletion of which led to reduced cellular growth or clonogenic survival in cells expressing HORMAD1. In addition to the translesion synthesis (TLS) polymerase POLH, we identified a HORMAD1-driven dependency upon additional TLS polymerases, namely POLK, REV1, REV3L and REV7. Our data confirms that out-of-context somatic expression of HORMAD1 can lead to genomic instability and reveals that HORMAD1 expression induces dependencies upon replication stress tolerance pathways, such as translesion synthesis. Our data also suggest that HORMAD1 expression could be a patient selection biomarker for agents targeting replication stress.
- Published
- 2022
226. Teaching PCR for Simultaneous Sensing of Gene Transcription and Downstream Metabolites by Cucurbit[8]uril-Mediated Intervention of Polymerase Activity
- Author
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Lancheng Wang, Mingjie Xu, Huimin Zhou, Kun Yan, Shiqi Duan, Dandan Xue, Youmei Wang, Bin Di, and Chi Hu
- Subjects
Macrocyclic Compounds ,Transcription, Genetic ,Ornithine Decarboxylase Inhibitors ,Imidazolidines ,Ornithine Decarboxylase ,Heterocyclic Compounds, 2-Ring ,Nucleotidyltransferases ,Polymerase Chain Reaction ,Analytical Chemistry - Abstract
The target of typical PCR analysis is restricted to nucleic acids. To this end, we report here a novel strategy to simultaneously detect genetic and metabolic markers using commercial PCR kits with cucurbit[8]urils (CB[8]) implemented to manipulate the activity of Taq DNA polymerase. CB[8] binds with the nonionic surfactants and displaces them from the polymerase surface, resulting in decreased enzyme activity. Meanwhile, the inhibited enzyme can be reversibly activated when spermine, the downstream metabolite of ornithine decarboxylase (ODC), is present in the sample, which competitively binds to CB[8] and recovers polymerase activity. CB[8] was implemented in conventional PCR kits not only to reduce false-positive results but also to extend the detection range of PCR technology. With this novel method to detect ODC in cell lysates containing both the nucleotides and intracellular metabolites, positive results were only observed in highly active HEK 293T cells, whereas silent cells treated with ODC inhibitor showed negative readouts, therefore providing a simple but elegant dual-modality PCR method for precision diagnosis.
- Published
- 2022
227. IL6 Induces mtDNA Leakage to Affect the Immune Escape of Endometrial Carcinoma via cGAS-STING
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Xue Zeng, Xiaosong Li, Yundong Zhang, Chaoxia Cao, and Qin Zhou
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Article Subject ,Interleukin-6 ,Immunology ,Membrane Proteins ,General Medicine ,DNA, Mitochondrial ,Nucleotidyltransferases ,B7-H1 Antigen ,Endometrial Neoplasms ,Mice ,Animals ,Humans ,Immunology and Allergy ,Female - Abstract
Endometrial carcinoma (EC) is a commonly diagnosed gynecological malignancy. Interleukin-6 (IL6) plays a critical role in modulating the progression of several types of tumors, including EC. However, the specific mechanism of IL6 in regulating EC progression has not been clearly elucidated. In this study, we performed a series of functional experiments to explore the potential mechanisms involved in IL6 function in the progression of EC. Here, we found that IL6 increased reactive oxygen species (ROS) generation by enhancing the NADPH oxidase (NOX) level and induced mtDNA leakage in EC cells, which further caused the activation of the downstream cGAS-STING signaling and increased production of extracellular vesicle (EV) production from EC cells. Besides, the activation of cGAS-STING signaling enhanced the expression of type I IFN and its downstream molecule PD-L1 through the TBK1-IRF3 pathway. Importantly, a high level mtDNA and PD-L1 were present in EVs derived from IL6-induced EC cells; these vesicles were shown to be able to induce T cell apoptosis. Finally, anti-PD-L1 treatment in mice showed that blockade of PD-L1 significantly reversed tumor immune escape mediated by IL6-induced EVs. Together, we provide evidence that IL6 induced mtDNA leakage to regulate the immune escape of EC cells. Our findings may provide a novel clue for the development of therapeutic targets for EC.
- Published
- 2022
228. One-pot platform for rapid detecting virus utilizing recombinase polymerase amplification and CRISPR/Cas12a
- Author
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Yifan Xiong, Gaihua Cao, Xiaolong Chen, Jun Yang, Meimei Shi, Yu Wang, Fuping Nie, Danqun Huo, and Changjun Hou
- Subjects
Recombinases ,Swine ,Animals ,General Medicine ,CRISPR-Cas Systems ,Real-Time Polymerase Chain Reaction ,African Swine Fever Virus ,Nucleic Acid Amplification Techniques ,Nucleotidyltransferases ,Sensitivity and Specificity ,Applied Microbiology and Biotechnology ,Biotechnology - Abstract
The livestock industry has been deeply affected by African swine fever virus (ASFV) and Capripoxvirus (CaPV), which caused an enormous economic damage. It is emergent to develop a reliable detection method. Here, we developed a rapid, ultra-sensitive, and one-pot DNA detection method combining recombinase polymerase amplification (RPA) and CRISPR/Cas12a for ASFV and CaPV, named one-pot-RPA-Cas12a (OpRCas) platform. It had the virtue of both RPA and CRISPR/Cas12a, such as high amplification efficiency, constant temperature reaction, and strict target selectivity, which made diagnosis simplified, accurate and easy to be operated without expensive equipment. Meanwhile, the reagents of RPA and CRISPR/Cas12a were added to the lid and bottom of tube in one go, which overcame the incompatibility of two reactions and aerosol contamination. To save cost, we only need a quarter of the amount of regular RPA per reaction which is enough to achieve clinical diagnosis. The OpRCas platform was 10 to 100 times more sensitive than qPCR; the limit of detection (LOD) was as low as 1.2 × 10
- Published
- 2022
229. cGAS-STING signaling in ischemic diseases
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Cai, Lei, Ying, Tan, Dan, Ni, Jinfu, Peng, and Guanghui, Yi
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Ischemia ,Biochemistry (medical) ,Clinical Biochemistry ,Humans ,Membrane Proteins ,General Medicine ,DNA, Mitochondrial ,Nucleotidyltransferases ,Biochemistry ,Signal Transduction - Abstract
Double stranded DNA (dsDNA) is known to act as a damage-associated molecular pattern (DAMP) that stimulates the body's innate immune response. In general, cyclicGMP-AMP(cGAMP)synthase(cGAS), a DNA sensor, detects these disease-causing DNA and activates the stimulator of interferon gene (STING), which in turn phosphorylates interferon regulatory factor 3 (IRF3), triggering the synthesis of type I interferon (IFN). During this process, the cGAS-STING pathway interacts with different modes of cell death, including autophagy, apoptosis, pyroptosis, and necroptosis. Importantly, cGAS might get stimulated by self-DNA, such as nuclear DNA (nuDNA) and mitochondrial DNA (mtDNA), which ensures a close association between the cGAS-STING signaling pathway and autoimmune responses. Following an ischemic attack, damaged or necrotic cells release large amounts of self-DNA that subsequently activate cGAS, resulting in a range of consequences related to an injury. The present study presents an overview of studies focused on cGAS-STING signaling and cell death, and summarizes the findings of this pathway with regard to ischemia or ischemia/reperfusion (I/R) in different organs of the body, including heart, brain, liver, kidney, and intestine.
- Published
- 2022
230. Cancer immunotherapy based on image-guided STING activation by nucleotide nanocomplex-decorated ultrasound microbubbles
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Xuefeng Li, Sina Khorsandi, Yifan Wang, Julien Santelli, Kristin Huntoon, Nhu Nguyen, Mingming Yang, DaeYong Lee, Yifei Lu, Ruoqi Gao, Betty Y. S. Kim, Caroline de Gracia Lux, Robert F. Mattrey, Wen Jiang, and Jacques Lux
- Subjects
Microbubbles ,Nucleotides ,Biomedical Engineering ,Antigen-Presenting Cells ,Membrane Proteins ,Bioengineering ,Condensed Matter Physics ,Nucleotidyltransferases ,Atomic and Molecular Physics, and Optics ,Nanostructures ,Mice ,Neoplasms ,Animals ,General Materials Science ,Immunotherapy ,Electrical and Electronic Engineering - Abstract
The cytosolic innate immune sensor cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway is crucial for priming adaptive antitumour immunity through antigen-presenting cells (APCs). Natural agonists, such as cyclic dinucleotides (CDNs), activate the cGAS-STING pathway, but their clinical translation is impeded by poor cytosolic entry and serum stability, low specificity and rapid tissue clearance. Here we developed an ultrasound (US)-guided cancer immunotherapy platform using nanocomplexes composed of 2'3'-cyclic guanosine monophosphate-adenosine monophosphate (cGAMP) electrostatically bound to biocompatible branched cationic biopolymers that are conjugated onto APC-targeting microbubbles (MBs). The nanocomplex-conjugated MBs engaged with APCs and efficiently delivered cGAMP into the cytosol via sonoporation, resulting in activation of cGAS-STING and downstream proinflammatory pathways that efficiently prime antigen-specific T cells. This bridging of innate and adaptive immunity inhibited tumour growth in both localized and metastatic murine cancer models. Our findings demonstrate that targeted local activation of STING in APCs under spatiotemporal US stimulation results in systemic antitumour immunity and improves the therapeutic efficacy of checkpoint blockade, thus paving the way towards novel image-guided strategies for targeted immunotherapy of cancer.
- Published
- 2022
231. UNC93B1 attenuates the cGAS–STING signaling pathway by targeting STING for autophagy–lysosome degradation
- Author
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Huifang Zhu, Rongzhao Zhang, Li Yi, Yan‐Dong Tang, and Chunfu Zheng
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Membrane Proteins ,Membrane Transport Proteins ,Interferon-beta ,Nucleotidyltransferases ,Immunity, Innate ,Mice ,HEK293 Cells ,Infectious Diseases ,Virology ,Autophagy ,Animals ,Humans ,Lysosomes ,Signal Transduction - Abstract
Stimulator of interferon genes (STING) is a pivotal innate immune adaptor, and its functions during DNA virus infections have been extensively documented. However, its homeostatic regulation is not well understood. Our study demonstrates that Unc-93 homolog B1 (UNC93B1) is a crucial checker for STING to prevent hyperactivation. Ectopic expression of UNC93B1 attenuates IFN-β promoter activity and the transcriptions of IFN-β, ISG54, and ISG56 genes. Moreover, UNC93B1 also blocks the IRF3 nuclear translocation induced by ectopic expression of both cyclic GMP-AMP synthase (cGAS) and STING and reduces the stability of STING by facilitating its autophagy-lysosome degradation, which can be reversed by lysosome inhibitors. Mechanistically, UNC93B1 interacts with STING and suppresses STING-activated downstream signaling by delivering STING to the lysosomes for degradation, depending on its trafficking capability. UNC93B1 knockout in human embryonic kidney 293T cells facilitates IFN-β promoter activity, IFN-β, ISG54, and ISG56 transcriptions, and IRF3 nuclear translocation induced by ectopic expression of cGAS and STING. Infected with herpes simplex virus-1 (HSV-1), UNC93B1 knockdown BJ cells or primary peritoneal macrophages from Unc93b1-deficient (Unc93b1
- Published
- 2022
232. Control of innate immunity by the <scp>cGAS‐STING</scp> pathway
- Author
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Mosallanejad, Kenta and Kagan, Jonathan C
- Subjects
Mammals ,Immunology ,Animals ,Membrane Proteins ,Immunology and Allergy ,DNA ,Interferons ,Cell Biology ,Nucleotidyltransferases ,Article ,Immunity, Innate - Abstract
Within the cytoplasm of mammalian cells is a protein called cyclic GMP-AMP synthase (cGAS), which acts to defend against infection and other threats to the host. cGAS operates in this manner through its ability to detect a molecular occurrence that should not exist in healthy cells - the existence of DNA in the cytosol. Upon DNA binding, cGAS synthesizes cyclic GMP-AMP (cGAMP), a cyclic dinucleotide that activates the endoplasmic reticulum-localized protein stimulator of interferon genes (STING). STING-mediated signaling culminates in host defensive responses typified by inflammatory cytokine and interferon expression, and the induction of autophagy. Studies over the past several years have established a consensus in the field of the enzymatic activities of cGAS in vitro, as it relates to DNA-induced production of cGAMP. However, much additional work is needed to understand the regulation of cGAS functions within cells, where multiple sources of DNA can create a problem of self and non-self discrimination. In this review, we provide an overview of how the cGAS-STING pathway mediates innate immune responses during infection and other cellular stresses. We then highlight recent progress in the understanding of the increasingly diverse ways in which this DNA-sensing machinery is regulated inside cells, including how cGAS remains inactive to host-derived DNA under conditions of homeostasis.
- Published
- 2022
233. The African swine fever virus protease pS273R inhibits DNA sensing cGAS-STING pathway by targeting IKKε
- Author
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Jia Luo, Jiajia Zhang, Jinghua Ni, Sen Jiang, Nengwen Xia, Yiwen Guo, Qi Shao, Qi Cao, Wanglong Zheng, Nanhua Chen, Quan Zhang, Hongjun Chen, Qing Chen, Hongfei Zhu, François Meurens, and Jianzhong Zhu
- Subjects
Microbiology (medical) ,Swine ,Immunology ,DNA ,African Swine Fever Virus ,Antiviral Agents ,Nucleotidyltransferases ,Microbiology ,Immunity, Innate ,I-kappa B Kinase ,Infectious Diseases ,Endopeptidases ,Animals ,Parasitology ,Peptide Hydrolases - Abstract
African swine fever virus (ASFV), a large and complex cytoplasmic double-stranded DNA virus, has developed multiple strategies to evade the antiviral innate immune responses. Cytosolic DNA arising from invading ASFV is mainly detected by the cyclic GMP-AMP synthase (cGAS) and then triggers a series of innate immune responses to prevent virus invasion. However, the immune escape mechanism of ASFV remains to be fully clarified. The pS273R of ASFV is a member of the SUMO-1-specific protease family and is crucial for valid virus replication. In this study, we identified pS273R as a suppressor of cGAS-STING pathway mediated type I interferon (IFN) production by ASFV genomic open reading frame screening. The pS273R was further confirmed as an inhibitor of IFN production as well as its downstream antiviral genes in cGAS-STING pathway. Mechanistically, pS273R greatly decreased the cGAS-STING signaling by targeting IKKε but not TBK1, and pS273R was found to disturb the interaction between IKKε and STING through its interaction with IKKε. Further, mutational analyses revealed that pS273R antagonized the cGAS-STING pathway by enzyme catalytic activity, which might affect the IKKε sumoylation state required for the interaction with STING. In summary, our results revealed for the first time that pS273R acts as an obvious negative regulator of cGAS-STING pathway by targeting IKKε via its enzymatic activity, which shows a new immune evasion mechanism of ASFV.
- Published
- 2022
234. ADP-Glucose Pyrophosphorylase Is Located in the Plastid in Developing Tomato Fruit
- Author
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Beckles, Diane M, Craig, Josephine, and Smith, Alison M
- Subjects
Plant Biology ,Biological Sciences ,Blotting ,Western ,Glucose-1-Phosphate Adenylyltransferase ,Solanum lycopersicum ,Nucleotidyltransferases ,Plastids ,Subcellular Fractions ,Agricultural and Veterinary Sciences ,Plant Biology & Botany ,Plant biology - Abstract
The subcellular location of activity and protein of ADP-glucose pyrophosphorylase (AGPase) in developing tomato (Lycopersicon esculentum) fruit was determined following a report that the enzyme might be present inside and outside the plastids in this organ. Plastids prepared from crude homogenates of columella and pericarp, the starch-accumulating tissues of developing fruit, contained 8% to 18% of the total activity of enzymes known to be confined to plastids, and 0.2% to 0.5% of the total activity of enzymes known to be confined to the cytosol. The proportion of the total activity of AGPase in the plastids was the same as that of the enzymes known to be confined to the plastid. When samples of plastid and total homogenate fractions were subjected to immunoblotting with an antiserum raised to AGPase, most or all of the protein detected was plastidial. Taken as a whole, these data provide strong evidence that AGPase is confined to the plastids in developing tomato fruit.
- Published
- 2001
235. A Cytosolic ADP-Glucose Pyrophosphorylase Is a Feature of Graminaceous Endosperms, But Not of Other Starch-Storing Organs
- Author
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Beckles, Diane M, Smith, Alison M, and Rees, Tom ap
- Subjects
Plant Biology ,Biological Sciences ,Cytosol ,Edible Grain ,Germination ,Glucose-1-Phosphate Adenylyltransferase ,Solanum lycopersicum ,Nucleotidyltransferases ,Plant Development ,Starch ,Trees ,Agricultural and Veterinary Sciences ,Plant Biology & Botany ,Plant biology - Abstract
The occurrence of an extra-plastidial isoform of ADP-glucose (Glc) pyrophosphorylase (AGPase) among starch-storing organs was investigated in two ways. First, the possibility that an extra-plastidial isoform arose during the domestication of cereals was studied by comparing the intracellular distribution of enzyme activity and protein in developing endosperm of noncultivated Hordeum species with that previously reported for cultivated barley (Hordeum vulgare). As in cultivated barley, the AGPase of H. vulgare subsp. spontaneum and Hordeum murinum endosperm is accounted for by a major extra-plastidial and a minor plastidial isoform. Second, the ratio of ADP-Glc to UDP-Glc was used as an indication of the intracellular location of the AGPase activity in a wide range of starch-synthesizing organs. The ratio is expected to be high in organs in which UDP-Glc and ADP-Glc are synthesized primarily in the cytosol, because the reactions catalyzed by AGPase and UDP-Glc pyrophosphorylase will be coupled and close to equilibrium. This study revealed that ADP-Glc contents and the ratio of ADP-Glc to UDP-Glc were higher in developing graminaceous endosperms than in any other starch-storing organs. Taken as a whole the results indicate that an extra-plastidial AGPase is important in ADP-Glc synthesis in graminaceous endosperms, but not in other starch-storing organs.
- Published
- 2001
236. Recombinase Polymerase Amplification Combined with Lateral Flow Dipstick Assay for the Rapid and Sensitive Detection of Pseudo-nitzschia multiseries .
- Author
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Yao Y, Luo N, Zong Y, Jia M, Rao Y, Huang H, and Jiang H
- Subjects
- Humans, Sensitivity and Specificity, Nucleotidyltransferases, DNA, Ribosomal, Nucleic Acid Amplification Techniques methods, Recombinases
- Abstract
The harmful algal bloom (HAB) species Pseudo-nitzschia multiseries is widely distributed worldwide and is known to produce the neurotoxin domoic acid, which harms marine wildlife and humans. Early detection and preventative measures are more critical than late management. However, the major challenge related to early detection is the accurate and sensitive detection of microalgae present in low abundance. Therefore, developing a sensitive and specific method that can rapidly detect P. multiseries is critical for expediting the monitoring and prediction of HABs. In this study, a novel assay method, recombinase polymerase amplification combined with lateral flow dipstick (RPA-LFD), is first developed for the detection of P. multiseries . To obtain the best test results, several important factors that affected the amplification effect were optimized. The internal transcribed spacer sequence of the nuclear ribosomal DNA from P. multiseries was selected as the target region. The results showed that the optimal amplification temperature and time for the recombinase polymerase amplification (RPA) of P. multiseries were 37 °C and 15 min. The RPA products could be visualized directly using the lateral flow dipstick after only 3 min. The RPA-LFD assay sensitivity for detection of recombinant plasmid DNA (1.9 × 10
0 pg/μL) was 100 times more sensitive than that of RPA, and the RPA-LFD assay sensitivity for detection of genomic DNA (2.0 × 102 pg/μL) was 10 times more sensitive than that of RPA. Its feasibility in the detection of environmental samples was also verified. In conclusion, these results indicated that the RPA-LFD detection of P. multiseries that was established in this study has high efficiency, sensitivity, specificity, and practicability. Management measures made based on information gained from early detection methods may be able to prevent certain blooms. The use of a highly sensitive approach for early warning detection of P. multiseries is essential to alleviate the harmful impacts of HABs on the environment, aquaculture, and human health.- Published
- 2024
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- View/download PDF
237. Cytoskeleton crosstalk: Casting stable actin bundles with dynamic microtubule molds.
- Author
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Haque F and Subramanian R
- Subjects
- Microtubules, Fungi, Nucleotidyltransferases, Actins, Cytoskeleton
- Abstract
Actin-microtubule crosstalk diversifies cytoskeletal networks. A new study provides insight into how the microtubule polymerase CKAP5 mediates actin-microtubule crosstalk. CKAP5 directs the assembly of stable actin bundles on dynamic microtubules; in turn, the actin bundles align growing microtubules along their length., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 Elsevier Inc. All rights reserved.)
- Published
- 2024
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- View/download PDF
238. Identification and Expression Analysis of the Nucleotidyl Transferase Protein (NTP) Family in Soybean ( Glycine max ) under Various Abiotic Stresses.
- Author
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Kang L, Li C, Qin A, Liu Z, Li X, Zeng L, Yu H, Wang Y, Song J, and Chen R
- Subjects
- Nucleotidyltransferases, Glycine max genetics, Cold-Shock Response, Nucleotides, RNA Nucleotidyltransferases, Arabidopsis, Arabidopsis Proteins
- Abstract
Nucleotidyl transferases (NTPs) are common transferases in eukaryotes and play a crucial role in nucleotide modifications at the 3' end of RNA. In plants, NTPs can regulate RNA stability by influencing 3' end modifications, which in turn affect plant growth, development, stress responses, and disease resistance. Although the functions of NTP family members have been extensively studied in Arabidopsis , rice, and maize, there is limited knowledge about NTP genes in soybeans. In this study, we identified 16 members of the NTP family in soybeans, including two subfamilies (G1 and G2) with distinct secondary structures, conserved motifs, and domain distributions at the protein level. Evolutionary analysis of genes in the NTP family across multiple species and gene collinearity analysis revealed a relatively conserved evolutionary pattern. Analysis of the tertiary structure of the proteins showed that NTPs have three conserved aspartic acids that bind together to form a possible active site. Tissue-specific expression analysis indicated that some NTP genes exhibit tissue-specific expression, likely due to their specific functions. Stress expression analysis showed significant differences in the expression levels of NTP genes under high salt, drought, and cold stress. Additionally, RNA-seq analysis of soybean plants subjected to salt and drought stress further confirmed the association of soybean NTP genes with abiotic stress responses. Subcellular localization experiments revealed that GmNTP2 and GmNTP14, which likely have similar functions to HESO1 and URT1, are located in the nucleus. These research findings provide a foundation for further investigations into the functions of NTP family genes in soybeans.
- Published
- 2024
- Full Text
- View/download PDF
239. Development of a Rapid Epstein-Barr Virus Detection System Based on Recombinase Polymerase Amplification and a Lateral Flow Assay.
- Author
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Sun Y, Tang D, Li N, Wang Y, Yang M, and Shen C
- Subjects
- Humans, Herpesvirus 4, Human genetics, Nucleotidyltransferases, Cell Line, Recombinases, Epstein-Barr Virus Infections diagnosis
- Abstract
The quality of cellular products used in biological research can directly impact the ability to obtain accurate results. Epstein-Barr virus (EBV) is a latent virus that spreads extensively worldwide, and cell lines used in experiments may carry EBV and pose an infection risk. The presence of EBV in a single cell line can contaminate other cell lines used in the same laboratory, affecting experimental results. We developed three EBV detection systems: (1) a polymerase chain reaction (PCR)-based detection system, (2) a recombinase polymerase amplification (RPA)-based detection system, and (3) a combined RPA-lateral flow assay (LFA) detection system. The minimum EBV detection limits were 1 × 10
3 copy numbers for the RPA-based and RPA-LFA systems and 1 × 104 copy numbers for the PCR-based system. Both the PCR and RPA detection systems were applied to 192 cell lines, and the results were consistent with those obtained by the EBV assay methods specified in the pharmaceutical industry standards of the People's Republic of China. A total of 10 EBV-positive cell lines were identified. The combined RPA-LFA system is simple to operate, allowing for rapid result visualization. This system can be implemented in laboratories and cell banks as part of a daily quality control strategy to ensure cell quality and experimental safety and may represent a potential new technique for the rapid detection of EBV in clinical samples.- Published
- 2024
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- View/download PDF
240. Rapid Detection of the Strawberry Foliar Nematode Aphelenchoides fragariae Using Recombinase Polymerase Amplification Assay with Lateral Flow Dipsticks.
- Author
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Subbotin SA
- Subjects
- Animals, Recombinases, Nucleotidyltransferases, DNA, Plant, Fragaria, Rhabditida, Tylenchida genetics
- Abstract
Rapid and reliable diagnostic methods for plant-parasitic nematodes are critical for facilitating the selection of effective control measures. A diagnostic recombinase polymerase amplification (RPA) assay for Aphelenchoides fragariae using a TwistAmp
® Basic Kit (TwistDx, Cambridge, UK) and AmplifyRP® Acceler8® Discovery Kit (Agdia, Elkhart, IN, USA) combined with lateral flow dipsticks (LF) has been developed. In this study, a LF-RPA assay was designed that targets the ITS rRNA gene of A. fragariae. This assay enables the specific detection of A. fragariae from crude nematode extracts without a DNA extraction step, and from DNA extracts of plant tissues infected with this nematode species. The LF-RPA assay showed reliable detection within 18-25 min with a sensitivity of 0.03 nematode per reaction tube for crude nematode extracts or 0.3 nematode per reaction tube using plant DNA extracts from 0.1 g of fresh leaves. The LF-RPA assay was developed and validated with a wide range of nematode and plant samples. Aphelenchoides fragariae was identified from seed samples in California. The LF-RPA assay has great potential for nematode diagnostics in the laboratory with minimal available equipment.- Published
- 2024
- Full Text
- View/download PDF
241. cGAS-STING, inflammasomes and pyroptosis: an overview of crosstalk mechanism of activation and regulation.
- Author
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Liu J, Zhou J, Luan Y, Li X, Meng X, Liao W, Tang J, and Wang Z
- Subjects
- Nucleotidyltransferases, Immunity, Innate, Caspase 1, Inflammasomes, Pyroptosis
- Abstract
Background: Intracellular DNA-sensing pathway cGAS-STING, inflammasomes and pyroptosis act as critical natural immune signaling axes for microbial infection, chronic inflammation, cancer progression and organ degeneration, but the mechanism and regulation of the crosstalk network remain unclear. Cellular stress disrupts mitochondrial homeostasis, facilitates the opening of mitochondrial permeability transition pore and the leakage of mitochondrial DNA to cell membrane, triggers inflammatory responses by activating cGAS-STING signaling, and subsequently induces inflammasomes activation and the onset of pyroptosis. Meanwhile, the inflammasome-associated protein caspase-1, Gasdermin D, the CARD domain of ASC and the potassium channel are involved in regulating cGAS-STING pathway. Importantly, this crosstalk network has a cascade amplification effect that exacerbates the immuno-inflammatory response, worsening the pathological process of inflammatory and autoimmune diseases. Given the importance of this crosstalk network of cGAS-STING, inflammasomes and pyroptosis in the regulation of innate immunity, it is emerging as a new avenue to explore the mechanisms of multiple disease pathogenesis. Therefore, efforts to define strategies to selectively modulate cGAS-STING, inflammasomes and pyroptosis in different disease settings have been or are ongoing. In this review, we will describe how this mechanistic understanding is driving possible therapeutics targeting this crosstalk network, focusing on the interacting or regulatory proteins, pathways, and a regulatory mitochondrial hub between cGAS-STING, inflammasomes, and pyroptosis., Short Conclusion: This review aims to provide insight into the critical roles and regulatory mechanisms of the crosstalk network of cGAS-STING, inflammasomes and pyroptosis, and to highlight some promising directions for future research and intervention., (© 2024. The Author(s).)
- Published
- 2024
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242. Real-time detection of Seneca Valley virus by one-tube RPA-CRISPR/Cas12a assay.
- Author
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Ma L, Zhu M, Meng Q, Wang Y, and Wang X
- Subjects
- Animals, Swine, CRISPR-Cas Systems, Nucleotidyltransferases, Recombinases, Picornaviridae
- Abstract
Introduction: Senecavirus A (SVA) is a highly contagious virus that causes vesicular disease in pigs. At present, laboratory detection methods, such as virus isolation and polymerase chain reaction (PCR), required precision instruments and qualified personnel, making them unsuitable for point-of-care tests (POCT). Fortunately, the emergence of CRISPR/Cas system has provided new opportunities for fast and efficient pathogen detection., Methods: This study successfully developed a precise and sensitive detection platform for diagnosing SVA by combining the CRISPR system with recombinase polymerase amplification (RPA)., Results: The minimum detection limit of the assay was 10 copies of the SVA genome. Meanwhile, the assay demonstrated high specificity. To validate the effectiveness of this system, we tested 85 swine clinical samples and found that the fluorescence method had a 100% coincidence rate compared to RT-qPCR., Discussion: Overall, the RPA-CRISPR/Cas12a assay established in our study is a highly effective method for detecting SVA and holds great potential for practical applications in the resource-limited settings., Competing Interests: Author QM was employed by the company Shanghai Kaiwosha Biotechnology Co., Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Ma, Zhu, Meng, Wang and Wang.)
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- 2024
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243. Macrophage extracellular traps promote tumor-like biologic behaviors of fibroblast-like synoviocytes through cGAS-mediated PI3K/Akt signaling pathway in patients with rheumatoid arthritis.
- Author
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Weng W, Liu Y, Hu Z, Li Z, Peng X, Wang M, Dong B, Zhong S, Jiang Y, and Pan Y
- Subjects
- Humans, Proto-Oncogene Proteins c-akt metabolism, Phosphatidylinositol 3-Kinases metabolism, Cell Proliferation, Signal Transduction, Nucleotidyltransferases, Fibroblasts, DNA metabolism, Cells, Cultured, Synoviocytes metabolism, Extracellular Traps metabolism, Arthritis, Rheumatoid pathology, Neoplasms pathology, Biological Products pharmacology
- Abstract
Rheumatoid arthritis is an autoimmune disease characterized by synovium hyperplasia and bone destruction. Macrophage extracellular traps are released from macrophages under various stimuli and may generate stable autoantigen-DNA complexes, as well as aggravate autoantibody generation and autoimmune responses. We aimed to investigate the role of macrophage extracellular traps on the biologic behaviors of rheumatoid arthritis fibroblast-like synoviocytes. Synovial tissues and fibroblast-like synoviocytes were obtained from patients with rheumatoid arthritis. Extracellular traps in synovium and synovial fluids were detected by immunofluorescence, immunohistochemistry, and SYTOX Green staining. Cell viability, migration, invasion, and cytokine expression of rheumatoid arthritis fibroblast-like synoviocytes were assessed by CCK-8, wound-healing assay, Transwell assays, and quantitative real-time polymerase chain reaction, respectively. RNA sequencing analysis was performed to explore the underlying mechanism, and Western blot was used to validate the active signaling pathways. We found that extracellular trap formation was abundant in rheumatoid arthritis and positively correlated to anti-CCP. Rheumatoid arthritis fibroblast-like synoviocytes stimulated with purified macrophage extracellular traps demonstrated the obvious promotion in tumor-like biologic behaviors. The DNA sensor cGAS in rheumatoid arthritis fibroblast-like synoviocytes was activated after macrophage extracellular trap stimuli. RNA sequencing revealed that differential genes were significantly enriched in the PI3K/Akt signaling pathway, and cGAS inhibitor RU.521 effectively reversed the promotion of tumor-like biologic behaviors in macrophage extracellular trap-treated rheumatoid arthritis fibroblast-like synoviocytes and downregulated the PI3K/Akt activation. In summary, our study demonstrates that macrophage extracellular traps promote the pathogenically biological behaviors of rheumatoid arthritis fibroblast-like synoviocytes through cGAS-mediated activation of the PI3K/Akt signaling pathway. These findings provide a novel insight into the pathogenesis of rheumatoid arthritis and the mechanisms of macrophages in modulating rheumatoid arthritis fibroblast-like synoviocyte tumor-like behaviors., Competing Interests: Conflict of interest statement. The authors have declared no conflicts of interest., (© The Author(s) 2023. Published by Oxford University Press on behalf of Society for Leukocyte Biology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
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- 2024
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244. Differential Poly(A) Tail Length Analysis Using Nanopore Sequencing.
- Author
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Dar SA, Malla S, Belair C, and Maragkakis M
- Subjects
- Nucleotidyltransferases, RNA, Messenger genetics, RNA, Messenger metabolism, Nucleotides, Poly A genetics, Poly A metabolism, Nanopore Sequencing
- Abstract
The poly(A) tail is a sequence of several adenosine nucleotides added to the 3' end of RNA molecules transcribed by polymerase II. The dynamics of poly(A) tail length play a significant role in regulating post-transcriptional gene expression by regulating the stability, translation, and decay of messenger RNAs. As a result, an accurate measurement of poly(A) tail length changes is important for understanding its regulatory function in different cellular contexts. Here, we outline a method for using nanopore sequencing and linear mixed models to analyze differences in poly(A) tail length across conditions., (© 2024. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.)
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- 2024
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245. Investigating the role of chlorogenic acids and coffee type in coffee-induced teeth discoloration.
- Author
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Kim S, Chung SH, Kim RJY, and Park YS
- Subjects
- Animals, Cattle, Humans, Chlorogenic Acid adverse effects, Beverages, Spectrophotometry, Nucleotidyltransferases, Color, Materials Testing, Tooth Discoloration
- Abstract
Objective: Coffee is one of the most popular beverages in the world, with millions of people consuming it every day. The effect of coffee on teeth discoloration has long been a concern for both coffee drinkers and dental professionals. To address this concern, this study aimed to investigate the role of chlorogenic acids (CGAs) and the type of coffee in coffee-induced teeth discoloration., Materials and Methods: High-performance liquid chromatography with a photodiode array detector was used to determine the CGA contents of instant coffee produced by five manufacturers (Starbucks, Dunkin' Donuts, Kanu, Ediya, Coffee Bean). A total of 180 bovine tooth specimens were immersed in the coffee samples for varying durations (3, 9, 24, 48, and 72 h), and the discoloration levels were measured using a spectrophotometer. A linear mixed-effects model analysis was used to determine the significance of L*, a*, and b* values in relation to the duration of coffee immersion and coffee type., Results: Both immersion time and coffee type had significant effects on tooth discoloration ( p < 0.001), with some types of coffee being more strongly associated with tooth discoloration than others. The amount of CGAs present in coffee was found to be positively correlated with the degree of discoloration ( p = 0.030)., Conclusions: Prolonged exposure to coffee can exacerbate teeth staining, and different types of coffee can cause varying degrees of discoloration. Furthermore, coffee with higher levels of CGAs may lead to greater tooth discoloration.
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- 2024
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246. Inhibition of ferroptosis by POLE2 in gastric cancer cells involves the activation of NRF2/GPX4 pathway.
- Author
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Jian H, Chen ZQ, Du H, Liao T, Sun YC, Ke D, and Yu Y
- Subjects
- Humans, Lentivirus, NF-E2-Related Factor 2, Nucleotidyltransferases, Protein Subunits, Ferroptosis, Stomach Neoplasms
- Abstract
Gastric cancer results in great cancer mortality worldwide, and inducing ferroptosis dramatically improves the malignant phenotypes of gastric cancer. DNA polymerase epsilon subunit 2 (POLE2) plays indispensable roles in tumorigenesis; however, its involvement and molecular basis in ferroptosis and gastric cancer are not clear. Human gastric cancer cells were infected with lentiviral vectors to knock down or overexpress POLE2, and cell ferroptosis was detected. To further validate the involvement of nuclear factor erythroid 2-related factor 2 (NRF2) and glutathione peroxidase 4 (GPX4), lentiviral vectors were used. POLE2 expression was elevated in human gastric cancer cells and tissues and closely correlated with clinicopathological features in gastric cancer patients. POLE2 knockdown was induced, while POLE2 overexpression inhibited ferroptosis of human gastric cancer cells, thereby modulating the malignant phenotypes of gastric cancer. Mechanistic studies revealed that POLE2 overexpression elevated NRF2 expression and activity and subsequently activated GPX4, which then prevented lipid peroxidation and ferroptosis in human gastric cancer cells. In contrast, either NRF2 or GPX4 silence significantly prevented POLE2 overexpression-mediated inductions of cell proliferation, migration, invasion and inhibition of ferroptosis. POLE2 overexpression inhibits ferroptosis in human gastric cancer cells through activating NRF2/GPX4 pathway, and inhibiting POLE2 may be a crucial strategy to treat gastric cancer., (© 2023 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.)
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- 2024
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247. A nuclear STING in the tail for canonical signalling.
- Author
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Minton K
- Subjects
- Humans, Signal Transduction, Nucleotidyltransferases
- Published
- 2024
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248. 3-N-butylphthalide attenuates neuroinflammation in rotenone-induced Parkinson's disease models via the cGAS-STING pathway.
- Author
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Liu Y, Duan R, Li P, Zhang B, and Liu Y
- Subjects
- Humans, Mice, Animals, Rotenone, NF-KappaB Inhibitor alpha, Neuroinflammatory Diseases, Reactive Oxygen Species, Quality of Life, DNA, Mitochondrial, Nucleotidyltransferases, Parkinson Disease drug therapy, Benzofurans
- Abstract
Neuroinflammation is crucial in the onset and progression of dopaminergic neuron loss in Parkinson's disease (PD). We aimed to determine whether 3-N-Butylphthalide (NBP) can protect against PD by inhibiting the cyclic GMP-AMP synthase (cGAS) - stimulator of interferon genes (STING) pathway and the inflammatory response of microglia. MitoSOX/MitoTracker/Hoechst staining was used to detect the levels of mitochondrial reactive oxygen species (ROS) in BV2 cells. Quantitative Real-Time Polymerase Chain Reaction was used to measure the levels of free cytoplasmic mitochondrial DNA (mtDNA) in BV2 cells and mouse brain tissues. Behavioral impairments were assessed using rotarod, T-maze, and balance beam tests. Dopaminergic neurons and microglia were observed using immunohistochemical staining. Expression levels of cGAS, STING, nuclear factor kappa-B (NfκB), phospho- NfκB (p-NfκB), inhibitor of NfκBα (IκBα), and phospho-IκBα (p-IκBα) proteins in the substantia nigra and striatum were detected using Western Blot. NBP decreased mitochondrial ROS levels in rotenone-treated BV2 cells. NBP alleviated behavioral impairments and protected against rotenone-induced microgliosis and damage to dopaminergic neurons in the substantia nigra and striatum of rotenone-induced PD mice. NBP decreased rotenone-induced mtDNA leakage and mitigated neuroinflammation by inhibiting cGAS-STING pathway activation. NBP exhibited a protective effect in rotenone-induced PD models by significantly inhibiting the cGAS-STING pathway. Moreover, NBP can alleviate neuroinflammation, and is a potential therapeutic drug for alleviating clinical symptoms and delaying the progression of PD. This study provided insights for the potential role of NBP in PD therapy, potentially mitigating neurodegeneration, and consequently improving the quality of life and lifespan of patients with PD. The limitations are that we have not confirmed the exact mechanism by which NBP decreases mtDNA leakage, and this study was unable to observe the actual clinical therapeutic effect, so further cohort studies are required for validation., Competing Interests: Declaration of conflicting interestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.
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- 2024
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249. The Role of the Toll-like Receptor 2 and the cGAS-STING Pathways in Breast Cancer: Friends or Foes?
- Author
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Cossu C, Di Lorenzo A, Fiorilla I, Todesco AM, Audrito V, and Conti L
- Subjects
- Humans, Female, Toll-Like Receptor 2, Breast, Drug Development, Nucleotidyltransferases, Tumor Microenvironment, Breast Neoplasms
- Abstract
Breast cancer stands as a primary malignancy among women, ranking second in global cancer-related deaths. Despite treatment advancements, many patients progress to metastatic stages, posing a significant therapeutic challenge. Current therapies primarily target cancer cells, overlooking their intricate interactions with the tumor microenvironment (TME) that fuel progression and treatment resistance. Dysregulated innate immunity in breast cancer triggers chronic inflammation, fostering cancer development and therapy resistance. Innate immune pattern recognition receptors (PRRs) have emerged as crucial regulators of the immune response as well as of several immune-mediated or cancer cell-intrinsic mechanisms that either inhibit or promote tumor progression. In particular, several studies showed that the Toll-like receptor 2 (TLR2) and the cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathways play a central role in breast cancer progression. In this review, we present a comprehensive overview of the role of TLR2 and STING in breast cancer, and we explore the potential to target these PRRs for drug development. This information will significantly impact the scientific discussion on the use of PRR agonists or inhibitors in cancer therapy, opening up new and promising avenues for breast cancer treatment.
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- 2023
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250. Bacillus amyloliquefaciens B10 Alleviates the Immunosuppressive Effects of Deoxynivalenol and Porcine Circovirus Type 2 Infection.
- Author
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Xiao H, Qin Z, Xu B, Long M, Wu Q, Guo X, Zhang H, Li Z, and Wu W
- Subjects
- Animals, Swine, Crops, Agricultural, Nucleotidyltransferases, Antiviral Agents, Bacillus amyloliquefaciens, Circovirus, Interferon Type I, Mycotoxins, Trichothecenes
- Abstract
As one of the most common mycotoxins, deoxynivalenol (DON) can contaminate a wide range of crops and foods. Porcine circovirus 2 (PCV2) is a kind of immunosuppressive virus, which can cause porcine circovirus associated disease (PCVD) in pig farms infected with PCV2. Pigs are extremely sensitive to DON, and PCV2-infected pig farms are often contaminated with DON. Our previous studies indicated that Bacillus amyloliquefaciens B10 (B10) has the potential to alleviate the toxicity of mycotoxins. The research was aimed at investigating the effects of Bacillus amyloliquefaciens B10 on the immunosuppressive effects caused by both DON and PCV2 infection. The results indicated that the expression of the PCV2 capsid protein CAP was significantly decreased after pretreatment with Bacillus amyloliquefaciens B10. Then, the effects of the Bacillus amyloliquefaciens B10 pretreatment on the type I interferon, antiviral protein and the antiviral signal pathway cGAS-STING was further investigated. The findings displayed that the expression of the type I interferon and antiviral protein were increased, while the IL-10 were decreased after pretreatment with Bacillus amyloliquefaciens B10. The inhibition of DON on the cGAS-STING signal pathway was relieved. Furthermore, it was found that this intervention effect was produced by inhibiting autophagy. In summary, Bacillus amyloliquefaciens B10 can mitigate the immunosuppressive effects of PCV2 and DON by inhibiting the production of autophagy.
- Published
- 2023
- Full Text
- View/download PDF
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