16 results on '"Jun-Dal, Kim"'
Search Results
2. Transcriptomic changes involved in the dedifferentiation of myofibroblasts derived from the lung of a patient with idiopathic pulmonary fibrosis
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Jun Ikari, Keita Ugai, Shuichi Matsuda, Akiyoshi Fukamizu, Hideki Mikami, Yoshitoshi Kasuya, Koichiro Tatsumi, Masahiko Hatano, Kenichi Suzuki, Jun-Dal Kim, and Kento Yoshioka
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0301 basic medicine ,Male ,Cancer Research ,Cell ,microRNA array ,Gene Expression ,Biochemistry ,Extracellular matrix ,Transcriptome ,03 medical and health sciences ,Idiopathic pulmonary fibrosis ,0302 clinical medicine ,transcriptome analysis ,Genetics ,medicine ,Humans ,Myofibroblasts ,Molecular Biology ,Lung ,Cells, Cultured ,Oncogene ,business.industry ,dedifferentiation ,Cell Differentiation ,Articles ,Azepines ,Middle Aged ,Triazoles ,respiratory system ,medicine.disease ,idiopathic pulmonary fibrosis ,Molecular medicine ,myofibroblast ,Actins ,Fibronectins ,respiratory tract diseases ,030104 developmental biology ,medicine.anatomical_structure ,Oncology ,030220 oncology & carcinogenesis ,Cancer research ,Molecular Medicine ,business ,Myofibroblast - Abstract
Idiopathic pulmonary fibrosis (IPF) is a fatal lung disease of unknown etiology. Under pathological conditions in lungs with IPF, myofibroblasts serve a key role in fibrogenesis via the accumulation of an excessive amount of extracellular matrix. To develop effective therapeutic interventions against IPF, studies have recently focused on how to dedifferentiate established myofibroblasts. The present study revealed that JQ1, an inhibitor of bromodomain and extra‑terminal proteins, markedly suppressed the expression levels of α‑smooth muscle actin and ED‑A‑fibronectin in myofibroblasts prepared from the lung of a patient with end‑stage IPF. Furthermore, these findings were supported by transcriptome analysis using RNA sequencing, in which differentially expressed genes (DEGs) downregulated by JQ1 treatment were significantly enriched in the fibrosis‑related signaling pathway. On the other hand, the upregulated DEGs in response to JQ1 treatment were significantly enriched in glutathione metabolism, which may affect the cell status of fibroblast/myofibroblast. To the best of our knowledge, this was the first study to comprehensively analyze transcriptome profiles associated with dedifferentiation of IPF myofibroblasts.
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- 2020
3. Cooperative action of APJ and α1A-adrenergic receptor in vascular smooth muscle cells induces vasoconstriction
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Mei Murao, Akiyoshi Fukamizu, Kenjiro Kimura, Yoshitoshi Kasuya, Chulwon Kwon, Katsumasa Nagano, Sadao Kimura, James T. Pearson, Nana Kishikawa, Jun-Dal Kim, Mikiyasu Shirai, Yi Ching Chen, Tatsuo Hashimoto, Hirotsugu Tsuchimochi, and Junji Ishida
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Agonist ,medicine.medical_specialty ,Vascular smooth muscle ,Adrenergic receptor ,medicine.drug_class ,Mice, Transgenic ,Vasodilation ,030204 cardiovascular system & hematology ,Biochemistry ,Muscle, Smooth, Vascular ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Receptors, Adrenergic, alpha-1 ,Internal medicine ,medicine ,Animals ,Humans ,Molecular Biology ,Phenylephrine ,030304 developmental biology ,Apelin receptor ,Apelin Receptors ,Mice, Inbred ICR ,0303 health sciences ,Chemistry ,General Medicine ,Apelin ,Endocrinology ,Vasoconstriction ,medicine.symptom ,medicine.drug - Abstract
The apelin receptor (APJ), a receptor for apelin and elabela/apela, induces vasodilation and vasoconstriction in blood vessels. However, the prolonged effects of increased APJ-mediated signalling, involving vasoconstriction, in smooth muscle cells have not been fully characterized. Here, we investigated the vasoactive effects of APJ gain of function under the control of the smooth muscle actin (SMA) gene promoter in mice. Transgenic overexpression of APJ (SMA-APJ) conferred sensitivity to blood pressure and vascular contraction induced by apelin administration in vivo. Interestingly, ex vivo experiments showed that apelin markedly increased the vasoconstriction of isolated aorta induced by noradrenaline (NA), an agonist for α- and β-adrenergic receptors, or phenylephrine, a specific agonist for α1-adrenergic receptor (α1-AR). In addition, intracellular calcium influx was augmented by apelin with NA in HEK293T cells expressing APJ and α1A-AR. To examine the cooperative action of APJ and α1A-AR in the regulation of vasoconstriction, we developed α1A-AR deficient mice using a genome-editing technique, and then established SMA-APJ/α1A-AR-KO mice. In the latter mouse line, aortic vasoconstriction induced by a specific agonist for α1A-AR, A-61603, were significantly less than in SMA-APJ mice. These results suggest that the APJ-enhanced response requires α1A-AR to contract vessels coordinately.
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- 2019
4. KDM5D-mediated H3K4 demethylation is required for sexually dimorphic gene expression in mouse embryonic fibroblasts
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Saori Tabara, Misaki Nakashima, Jun-Dal Kim, Weizhe Lu, Chulwon Kwon, Akiyoshi Fukamizu, and Hayase Mizukami
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Male ,RNA-Seq ,Biology ,Methylation ,Biochemistry ,Histones ,Transcriptome ,Mice ,03 medical and health sciences ,Gene expression ,Animals ,Molecular Biology ,Gene ,030304 developmental biology ,Histone Demethylases ,Genetics ,Sex Characteristics ,0303 health sciences ,030302 biochemistry & molecular biology ,General Medicine ,Fibroblasts ,Embryo, Mammalian ,Gene expression profiling ,Testis determining factor ,Gene Expression Regulation ,Gene Knockdown Techniques ,biology.protein ,Demethylase ,Female - Abstract
Males and females share the same genetic code, but gene expression profile often displays differences between two sexes. Mouse embryonic fibroblasts (MEFs) have been used to experiment as a useful tool to test gene function. They have also been characterized by gender-based differences in expressed genes such as Y-linked Sry or X-linked Hprt. However, there is no report on sex differences in global gene expression. Here, using the next-generation RNA sequencing, we compared the comprehensive transcriptome of MEFs derived from two sexes. In comparison with the female group, the male group up-regulated 27 differentially expressed genes (DEGs), in which a male-specific histone demethylase KDM5D gene is included, and 7 DEGs were down-regulated. Based on the results by searching the ENCODE analysis, it was shown that the expression of 15 genes identified is potentially regulated by the methylation of H3K4me1 or H3K4me3. Interestingly, we demonstrated that both of H3K4 methylation are induced by knocking down KDM5D, which causes changes in patterns of eight DEGs found in male MEFs. Collectively, these data not only suggest an importance of KDM5D-mediated demethylation of H3K4 involved in the sexually dimorphic gene expression in male MEFs, but also may provide information regarding sex-dependent changes in gene expression when MEFs are used for experiments.
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- 2018
5. Starvation-induced transcription factor CREBH negatively governs body growth by controlling GH signaling
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Hitoshi Shimano, Satoru Takahashi, Kae Kumagai, Masaya Araki, Takafumi Miyamoto, Yasunari Yamada, Takashi Matsuzaka, Morichika Konishi, Motohiro Sekiya, Nobuyuki Itoh, Yuhei Mizunoe, Seiya Mizuno, Hirohito Sone, Jun-Dal Kim, Yoshimi Nakagawa, Hiroshi Ohno, Kazuya Matsuo, and Song-iee Han
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0301 basic medicine ,Male ,medicine.medical_specialty ,FGF21 ,medicine.medical_treatment ,Transgene ,Mice, Transgenic ,Growth hormone receptor ,Biochemistry ,Body Mass Index ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Internal medicine ,Genetics ,medicine ,Animals ,IGFBP1 ,Cyclic AMP Response Element-Binding Protein ,Molecular Biology ,Transcription factor ,Mice, Knockout ,Messenger RNA ,Chemistry ,Growth factor ,Binding protein ,Gene Expression Regulation, Developmental ,Fibroblast Growth Factors ,Mice, Inbred C57BL ,030104 developmental biology ,Endocrinology ,Liver ,Growth Hormone ,Body Composition ,Female ,030217 neurology & neurosurgery ,Biotechnology ,Signal Transduction - Abstract
cAMP responsive element-binding protein H (CREBH) is a hepatic transcription factor to be activated during fasting. We generated CREBH knock-in flox mice, and then generated liver-specific CREBH transgenic (CREBH L-Tg) mice in an active form. CREBH L-Tg mice showed a delay in growth in the postnatal stage. Plasma growth hormone (GH) levels were significantly increased in CREBH L-Tg mice, but plasma insulin-like growth factor 1 (IGF1) levels were significantly decreased, indicating GH resistance. In addition, CREBH overexpression significantly increased hepatic mRNA and plasma levels of FGF21, which is thought to be as one of the causes of growth delay. However, the additional ablation of FGF21 in CREBH L-Tg mice could not correct GH resistance at all. CREBH L-Tg mice sustained GH receptor (GHR) reduction and the increase of IGF binding protein 1 (IGFBP1) in the liver regardless of FGF21. As GHR is a first step in GH signaling, the reduction of GHR leads to impairment of GH signaling. These data suggest that CREBH negatively regulates growth in the postnatal growth stage via various pathways as an abundant energy response by antagonizing GH signaling.
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- 2021
6. Transcriptomic Evaluation of Pulmonary Fibrosis-Related Genes: Utilization of Transgenic Mice with Modifying p38 Signal in the Lungs
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Kazuya Murata, Jun-Dal Kim, Junji Ishida, Koichiro Tatsumi, Kanako Nakamura, Kana Namiki, Tatsuhiko Sudo, Masahiko Hatano, Shuichi Matsuda, Tomoyuki Kuwaki, Akiyoshi Fukamizu, Fumihiro Sugiyama, Yuji Matsuo, and Yoshitoshi Kasuya
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Male ,0301 basic medicine ,Pulmonary compliance ,p38 Mitogen-Activated Protein Kinases ,lcsh:Chemistry ,Transcriptome ,Mice ,Idiopathic pulmonary fibrosis ,chemistry.chemical_compound ,0302 clinical medicine ,Fibrosis ,Pulmonary fibrosis ,lcsh:QH301-705.5 ,Lung ,Spectroscopy ,RNA sequencing ,General Medicine ,respiratory system ,idiopathic pulmonary fibrosis ,Computer Science Applications ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Female ,Collagen ,MAP Kinase Signaling System ,Mice, Transgenic ,Bleomycin ,Article ,Catalysis ,Inorganic Chemistry ,03 medical and health sciences ,Immune system ,medicine ,Animals ,Physical and Theoretical Chemistry ,Molecular Biology ,business.industry ,Organic Chemistry ,p38 mitogen-activated protein kinase ,alveolar epithelial type II cells ,medicine.disease ,respiratory tract diseases ,Mice, Inbred C57BL ,030104 developmental biology ,lcsh:Biology (General) ,lcsh:QD1-999 ,chemistry ,Alveolar Epithelial Cells ,Cancer research ,bleomycin-induced pulmonary fibrosis ,business - Abstract
Idiopathic pulmonary fibrosis (IPF) is a progressive fibrosing lung disease that is caused by the dysregulation of alveolar epithelial type II cells (AEC II). The mechanisms involved in the progression of IPF remain incompletely understood, although the immune response accompanied by p38 mitogen-activated protein kinase (MAPK) activation may contribute to some of them. This study aimed to examine the association of p38 activity in the lungs with bleomycin (BLM)-induced pulmonary fibrosis and its transcriptomic profiling. Accordingly, we evaluated BLM-induced pulmonary fibrosis during an active fibrosis phase in three genotypes of mice carrying stepwise variations in intrinsic p38 activity in the AEC II and performed RNA sequencing of their lungs. Stepwise elevation of p38 signaling in the lungs of the three genotypes was correlated with increased severity of BLM-induced pulmonary fibrosis exhibiting reduced static compliance and higher collagen content. Transcriptome analysis of these lung samples also showed that the enhanced p38 signaling in the lungs was associated with increased transcription of the genes driving the p38 MAPK pathway and differentially expressed genes elicited by BLM, including those related to fibrosis as well as the immune system. Our findings underscore the significance of p38 MAPK in the progression of pulmonary fibrosis.
- Published
- 2020
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7. PRMT1 Deficiency in Mouse Juvenile Heart Induces Dilated Cardiomyopathy and Reveals Cryptic Alternative Splicing Products
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Junji Ishida, Weizhe Lu, Kana Nishikata, Jun-Dal Kim, Kazuya Murata, Akiyoshi Fukamizu, Misuzu Hashimoto, Natsumi Ono, Masafumi Muratani, and Shizufumi Ebihara
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0301 basic medicine ,Gene isoform ,Messenger RNA ,Multidisciplinary ,Arginine ,Alternative splicing ,Skeletal muscle ,Cell Biology ,Methylation ,Biology ,Article ,Cell biology ,03 medical and health sciences ,030104 developmental biology ,medicine.anatomical_structure ,Ubiquitin ,Genetics ,medicine ,biology.protein ,lcsh:Q ,lcsh:Science ,Molecular Biology ,Gene ,Developmental Biology - Abstract
Summary Protein arginine methyltransferase 1 (PRMT1) catalyzes the asymmetric dimethylation of arginine residues in proteins and methylation of various RNA-binding proteins and is associated with alternative splicing in vitro. Although PRMT1 has essential in vivo roles in embryonic development, CNS development, and skeletal muscle regeneration, the functional importance of PRMT1 in the heart remains to be elucidated. Here, we report that juvenile cardiomyocyte-specific PRMT1-deficient mice develop severe dilated cardiomyopathy and exhibit aberrant cardiac alternative splicing. Furthermore, we identified previously undefined cardiac alternative splicing isoforms of four genes (Asb2, Fbxo40, Nrap, and Eif4a2) in PRMT1-cKO mice and revealed that eIF4A2 protein isoforms translated from alternatively spliced mRNA were differentially ubiquitinated and degraded by the ubiquitin-proteasome system. These findings highlight the essential roles of PRMT1 in cardiac homeostasis and alternative splicing regulation., Graphical Abstract, Highlights • PRMT1 deficiency in cardiomyocytes causes dilated cardiomyopathy in juvenile mice • PRMT1-deficient heart shows abnormal alternative splicing patterns • Previously undefined cardiac splicing events are revealed by transcriptome analysis • eIF4A2 isoforms are differentially ubiquitinated and degraded, Genetics; Molecular Biology; Cell Biology; Developmental Biology
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- 2018
8. Emerging impacts of biological methylation on genetic information
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Akiyoshi Fukamizu, Jun-Dal Kim, and Koichiro Kako
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Methyltransferase ,Central dogma of molecular biology ,Computational biology ,Biology ,Arginine ,Methylation ,Biochemistry ,03 medical and health sciences ,chemistry.chemical_compound ,Animals ,Humans ,Molecular Biology ,030304 developmental biology ,Whole genome sequencing ,0303 health sciences ,Lysine ,030302 biochemistry & molecular biology ,Proteins ,DNA ,Methyltransferases ,General Medicine ,DNA Methylation ,Ribosomal RNA ,Demethylation ,chemistry ,Protein Biosynthesis ,DNA methylation ,Transfer RNA ,RNA - Abstract
The central dogma of molecular biology explains the fundamental flow of genetic information for life. Although genome sequence (DNA) itself is a static chemical signature, it includes multiple layers of information composed of mRNA, tRNA, rRNA and small RNAs, all of which are involved in protein synthesis and is passing from parents to offspring via DNA. Methylation is a biologically important modification, because DNA, RNAs and proteins, components of the central dogma, are methylated by a set of methyltransferases. Recent works focused on understanding a variety of biological methylation have shed light on new regulation of cellular functions. In this review, we briefly discuss some of those recent findings of methylation, including DNA, RNAs and proteins.
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- 2018
9. Pathophysiological Roles of Stress-Activated Protein Kinases in Pulmonary Fibrosis
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Masahiko Hatano, Shuichi Matsuda, Koichiro Tatsumi, Jun-Dal Kim, and Yoshitoshi Kasuya
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MAP Kinase Kinase 4 ,MAP Kinase Signaling System ,QH301-705.5 ,p38 mitogen-activated protein kinases ,Connective tissue ,Review ,p38 MAPK ,p38 Mitogen-Activated Protein Kinases ,c-Jun NH2-terminal kinase ,Catalysis ,Inorganic Chemistry ,Idiopathic pulmonary fibrosis ,Fibrosis ,Pulmonary fibrosis ,medicine ,Humans ,Biology (General) ,Physical and Theoretical Chemistry ,Protein kinase A ,Lung ,QD1-999 ,Molecular Biology ,Spectroscopy ,Kinase ,business.industry ,Macrophages ,Organic Chemistry ,JNK Mitogen-Activated Protein Kinases ,stress-activated protein kinases ,General Medicine ,Fibroblasts ,idiopathic pulmonary fibrosis ,medicine.disease ,Computer Science Applications ,Chemistry ,medicine.anatomical_structure ,Cancer research ,Blood Vessels ,business - Abstract
Idiopathic pulmonary fibrosis (IPF) is one of the most symptomatic progressive fibrotic lung diseases, in which patients have an extremely poor prognosis. Therefore, understanding the precise molecular mechanisms underlying pulmonary fibrosis is necessary for the development of new therapeutic options. Stress-activated protein kinases (SAPKs), c-Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (p38) are ubiquitously expressed in various types of cells and activated in response to cellular environmental stresses, including inflammatory and apoptotic stimuli. Type II alveolar epithelial cells, fibroblasts, and macrophages are known to participate in the progression of pulmonary fibrosis. SAPKs can control fibrogenesis by regulating the cellular processes and molecular functions in various types of lung cells (including cells of the epithelium, interstitial connective tissue, blood vessels, and hematopoietic and lymphoid tissue), all aspects of which remain to be elucidated. We recently reported that the stepwise elevation of intrinsic p38 signaling in the lungs is correlated with a worsening severity of bleomycin-induced fibrosis, indicating an importance of this pathway in the progression of pulmonary fibrosis. In addition, a transcriptome analysis of RNA-sequencing data from this unique model demonstrated that several lines of mechanisms are involved in the pathogenesis of pulmonary fibrosis, which provides a basis for further studies. Here, we review the accumulating evidence for the spatial and temporal roles of SAPKs in pulmonary fibrosis.
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- 2021
10. The N-terminal sequence of murine PRMT5 variant 2 is required for Hsp70 interaction and CHIP ligase-mediated degradation
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Weizhe Lu, Jun-Dal Kim, Hayase Mizukami, Saori Tabara, Keiji Kimura, Chulwon Kwon, and Akiyoshi Fukamizu
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0301 basic medicine ,Male ,Protein-Arginine N-Methyltransferases ,Ubiquitin-Protein Ligases ,Biophysics ,Protein degradation ,Biochemistry ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Heat shock protein ,Histone H2A ,Animals ,Humans ,HSP70 Heat-Shock Proteins ,Molecular Biology ,Cells, Cultured ,Adaptor Proteins, Signal Transducing ,chemistry.chemical_classification ,DNA ligase ,Chemistry ,Protein arginine methyltransferase 5 ,Cell Biology ,Methylation ,Cell biology ,Hsp70 ,Mice, Inbred C57BL ,030104 developmental biology ,HEK293 Cells ,Proteasome ,030220 oncology & carcinogenesis ,Female - Abstract
Protein arginine methyltransferase PRMT5 synthesizes the symmetric dimethylarginine in nuclear and cytoplasmic proteins such as histone H2A, H4 and several non-histone proteins that are required for a variety of biological processes. Currently, two splice variants (v1 and v2) of murine PRMT5 have been deposited in the NCBI sequence database, in which PRMT5-v1 and -v2 contain different 33 and 16 amino acids at the N-terminal sequences, respectively. Here we showed that murine PRMT5-v1 is stable, but PRMT5-v2 is constantly degraded through both the ubiquitin proteasome system (UPS) and the autophagic-lysosomal pathway (ALP) in an N-terminal sequence-dependent manner. Furthermore, inhibition of UPS and ALP elevated the stability of PRMT5-v2 that made it localized in the nucleus and the cytoplasm. In addition, PRMT5-v2 exhibited the enzyme activity to catalyze histone H2A and H4 methylation. Notably, we found that the heat shock protein (Hsp) 70 specially recognizes the N-terminal sequence of PRMT5-v2 and the carboxyl terminus of Hsp70-interacting protein (CHIP) is required for poly-ubiquitination and the degradation of PRMT5-v2. These results suggest that Hsp70/CHIP chaperone-mediated protein degradation system is crucial in the regulation of PRMT5-v2 turnover, which has the potential to balance the symmetrical arginine dimethylation in cells.
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- 2019
11. Protein arginine methyltransferase 7 has a novel homodimer-like structure formed by tandem repeats
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Morio Hasegawa, Sachiko Toma-Fukai, Toshiyuki Shimizu, Akiyoshi Fukamizu, and Jun-Dal Kim
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Models, Molecular ,Protein Folding ,Protein-Arginine N-Methyltransferases ,Arginine ,Biophysics ,Crystallography, X-Ray ,Biochemistry ,Protein Structure, Secondary ,Catalysis ,Tandem repeat ,Protein arginine methyltransferase ,Structural Biology ,Catalytic Domain ,Scattering, Small Angle ,Genetics ,Animals ,Transferase ,Caenorhabditis elegans ,Caenorhabditis elegans Proteins ,Molecular Biology ,X-ray crystallography ,chemistry.chemical_classification ,Cofactor binding ,Binding Sites ,biology ,Chemistry ,Mutagenesis ,Cell Biology ,biology.organism_classification ,Protein Structure, Tertiary ,Enzyme ,Tandem Repeat Sequences ,Mutation ,Protein Multimerization ,S-adenosyl-l-homocysteine ,Protein Binding - Abstract
Protein arginine methyltransferase 7 (PRMT7) is a member of a family of enzymes that catalyze the transfer of methyl groups from S-adenosyl-l-methionine to nitrogen atoms on arginine residues. Here, we describe the crystal structure of Caenorhabditis elegans PRMT7 in complex with its reaction product S-adenosyl-l-homocysteine. The structural data indicated that PRMT7 harbors two tandem repeated PRMT core domains that form a novel homodimer-like structure. S-adenosyl-l-homocysteine bound to the N-terminal catalytic site only; the C-terminal catalytic site is occupied by a loop that inhibits cofactor binding. Mutagenesis demonstrated that only the N-terminal catalytic site of PRMT7 is responsible for cofactor binding.
- Published
- 2014
12. Truncated Cables1 causes agenesis of the corpus callosum in mice
- Author
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Dinh T H Tra, Akiyoshi Fukamizu, Atsushi Mizobuchi, Junji Ishida, Fumihiro Sugiyama, Yoichi Matsuda, Saori Mizuno-Iijima, Ken-ichi Yagami, Hiroyoshi Iseki, Jun-Dal Kim, Satoshi Kunita, and Seiya Mizuno
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Genetically modified mouse ,Transgene ,Mutant ,Mice, Transgenic ,Biology ,Corpus callosum ,Pathology and Forensic Medicine ,Mice ,Exon ,Cyclins ,Gene expression ,medicine ,Animals ,RNA, Messenger ,Insertion ,Agenesis of the corpus callosum ,Molecular Biology ,Genetic Association Studies ,In Situ Hybridization, Fluorescence ,Mice, Knockout ,Genetics ,Mice, Inbred BALB C ,Mice, Inbred ICR ,Homozygote ,Exons ,Cell Biology ,Phosphoproteins ,medicine.disease ,Molecular biology ,Mice, Inbred C57BL ,Mutagenesis, Insertional ,Agenesis of Corpus Callosum ,Carrier Proteins - Abstract
Agenesis of the corpus callosum (ACC) is a congenital abnormality of the brain structure. More than 60 genes are known to be involved in corpus callosum development. However, the molecular mechanisms underlying ACC are not fully understood. Previously, we produced a novel transgenic mouse strain, TAS, carrying genes of the tetracycline-inducible expression system that are not involved in brain development, and inherited ACC was observed in the brains of all homozygous TAS mice. Although ACC was probably induced by transgene insertion mutation, the causative gene and the molecular mechanism of its pathogenesis remain unclear. Here, we first performed interphase three-color fluorescence in situ hybridization (FISH) analysis to determine the genomic insertion site. Transgenes were inserted into chromosome 18 ∼12.0 Mb from the centromere. Gene expression analysis and genomic PCR walking showed that the genomic region containing exon 4 of Cables1 was deleted by transgene insertion and the other exons of Cables1 were intact. The mutant allele was designated as Cables1(TAS). Interestingly, Cables1(TAS) mRNA consisted of exons 1-3 of Cables1 and part of the transgene that encoded a novel truncated Cables1 protein. Homozygous TAS mice exhibited mRNA expression of Cables1(TAS) in the fetal cerebrum, but not that of wild-type Cables1. To investigate whether a dominant negative effect of Cables1(TAS) or complete loss of function of Cables1 gives rise to ACC, we produced Cables1-null mutant mice. ACC was not observed in Cables1-null mutant mice, suggesting that a dominant negative effect of Cables1(TAS) impairs callosal formation. Moreover, ACC frequency in Cables1(+/TAS) mice was significantly lower than that in Cables1(-/TAS) mice, indicating that wild-type Cables1 interfered with the dominant negative effect of Cables1(TAS). This study indicated that truncated Cables1 causes ACC and wild-type Cables1 contributes to callosal formation.
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- 2014
13. Enhanced histamine production through the induction of histidine decarboxylase expression by phorbol ester in Jurkat cells
- Author
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Yusuke Nagashima, Akiyoshi Fukamizu, Jun-Dal Kim, and Koichiro Kako
- Subjects
Cancer Research ,matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight tandem mass spectrometry ,Histidine Decarboxylase ,Biochemistry ,Jurkat cells ,Jurkat Cells ,chemistry.chemical_compound ,Genetics ,Humans ,RNA, Messenger ,Molecular Biology ,Chromatography, High Pressure Liquid ,Histamine Production ,Articles ,histamine ,Histidine decarboxylase ,Molecular biology ,Up-Regulation ,Oncology ,chemistry ,ultra-high performance liquid chromatography ,Apoptosis ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Tetradecanoylphorbol Acetate ,Quinolines ,Phorbol ,Molecular Medicine ,Histamine ,Intracellular ,phorbol 12-myristate 13-acetate - Abstract
Histamine (HA), a mediator of inflammation, type I allergic responses and neurotransmission, is synthesized from L-histidine, the reaction of which is catalyzed by histidine decarboxylase (HDC). HDC has been reported to be induced by various stimuli, not only in mast cells and basophils, but also in T lymphocytes and macrophages. Although its mRNA has been shown to be increased in Jurkat cells when treated with phorbol 12-myristate 13-acetate (TPA), little is known concerning the induced production of HA by HDC. The present study quantified the trace amounts of intracellular HA using ultra-high liquid chromatography in combination with the 6-aminoquinoline carbamate-derivatization technique. To test whether the cellular level of HA is elevated by the induction of HDC in Jurkat cells treated with TPA, the peak corresponding to authentic HA in the cell lysate was fractioned and its molecular weight determined by matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometry. The results of this study show that the HA level is increased by the induction of HDC expression by TPA in Jurkat cells. Therefore, this method is useful in elucidating the physiological significance of HA production.
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- 2012
14. Mechanism for p38α-mediated Experimental Autoimmune Encephalomyelitis
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Hirofumi Matsunaga, Kana Namiki, Yasunori Tokunaga, Kensuke Tanaka, Yoshitoshi Kasuya, Kazuya Murata, Akiyoshi Fukamizu, Akira Sakairi, Jun-Dal Kim, Manabu Shigetomi, Shibakawa Nobuhiko, Tatsuhiko Sudo, Sadao Kimura, Yukinori Wada, Naoki Tokuhara, Junji Ishida, Motohisa Shimizu, Kento Yoshioka, and Masahiko Hagihara
- Subjects
Male ,Encephalomyelitis, Autoimmune, Experimental ,RNA Stability ,p38 mitogen-activated protein kinases ,Encephalomyelitis ,Electrophoretic Mobility Shift Assay ,Biology ,Biochemistry ,Mitogen-Activated Protein Kinase 14 ,Pathogenesis ,Mice ,Western blot ,medicine ,Animals ,Electrophoretic mobility shift assay ,Enzyme Inhibitors ,Promoter Regions, Genetic ,Molecular Biology ,Gene knockout ,medicine.diagnostic_test ,Interleukin-17 ,Experimental autoimmune encephalomyelitis ,Cell Biology ,medicine.disease ,Molecular biology ,Female ,Interleukin 17 ,Signal Transduction - Abstract
One of the mitogen-activated protein kinases, p38, has been found to play a crucial role in various inflammatory responses. In this study, we analyzed the roles of p38α in multiple sclerosis, using an animal model, experimental autoimmune encephalomyelitis (EAE). p38α+/− mice (p38α−/− showed embryonic lethality) showed less severe neurological signs than WT mice. Adoptive transfer of lymph node cells (LNC) from sensitized WT mice with MOG(35–55) to naive WT-induced EAE was much more severe compared with the case using LNC from sensitized p38α+/− mice. Comprehensive analysis of cytokines from MOG(35–55)-challenged LNC by Western blot array revealed that production of IL-17 was significantly reduced by a single copy disruption of the p38α gene or a p38 inhibitor. Likewise, by a luciferase reporter assay, an electrophoresis mobility shift assay, and characterization of the relationship between p38 activity and IL-17 mRNA expression, we confirmed that p38 positively regulates transcription of the Il17 gene. Furthermore, oral administration of a highly specific p38α inhibitor (UR-5269) to WT mice at the onset of EAE markedly suppressed the progression of EAE compared with a vehicle group. These results suggest that p38α participates in the pathogenesis of EAE through IL-17 induction. Background: p38 signaling pathway plays a key role in inflammatory diseases. Results: A single copy disruption of the p38α gene or a p38α inhibitor markedly reduced the pathogenesis of EAE by decreasing IL-17 production. Conclusion: p38α regulates the pathogenesis of EAE through transcriptional regulation of IL-17 production. Significance: Anti-p38α strategy achieves therapeutic benefit for the treatment of multiple sclerosis.
- Published
- 2012
15. Detection of choline and phosphatidic acid (PA) catalyzed by phospholipase D (PLD) using MALDI-QIT-TOF/MS with 9-aminoacridine matrix
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Akiyoshi Fukamizu, Motoki Matsui, Kyung-Eui Park, Hiroaki Daitoku, Jun-Dal Kim, Yusuke Nagashima, Gwi Gun Park, and Koichiro Kako
- Subjects
Phosphatidic Acids ,Applied Microbiology and Biotechnology ,Biochemistry ,Analytical Chemistry ,Choline ,chemistry.chemical_compound ,9-Aminoacridine ,Limit of Detection ,Phosphatidylcholine ,medicine ,Phospholipase D ,Phospholipase D activity ,Humans ,Molecular Biology ,Enzyme Assays ,Chromatography ,Chemistry ,PLD2 ,Hydrolysis ,Organic Chemistry ,General Medicine ,Phosphatidic acid ,Aminacrine ,HEK293 Cells ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Biocatalysis ,lipids (amino acids, peptides, and proteins) ,Acetylcholine ,Biotechnology ,medicine.drug - Abstract
Phospholipase D (PLD) catalyzes the hydrolysis of phosphatidylcholine (PC), the most abundant phospholipids of plasma membrane, resulting in the production of choline and phosphatidic acid (PA). Choline is a precursor of the neurotransmitter acetylcholine, whereas PA functions as an intracellular lipid mediator of diverse biological functions. For assessing PLD activity in vitro, PLD-derived choline has been often analyzed with radioactive or non-radioactive methods. In this study, we have developed a new method for detecting choline and PA with MALDI-QIT-TOF/MS by using 9-aminoacridine as a matrix. The standard calibration curves showed that choline and PA could be detected with linearity over the range from 0.05 and 1 pmol, respectively. Importantly, this method enables the concomitant detection of choline and PA as a reaction product of PC hydrolysis by PLD2 proteins. Thus, our simple and direct method would be useful to characterize the enzymatic properties of PLD, thereby providing insight into mechanisms of PLD activation.
- Published
- 2014
16. Angiodysplasia in embryo lacking protein arginine methyltransferase 1 in vascular endothelial cells
- Author
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Tomohiro Ishimaru, Kanako Hara, Hayase Mizukami, Ken-ichi Yagami, Akiyoshi Fukamizu, Jun-Dal Kim, Fumihiro Sugiyama, Junji Ishida, and Misuzu Hashimoto
- Subjects
0301 basic medicine ,Protein-Arginine N-Methyltransferases ,Angiogenesis ,Mice, Transgenic ,Biology ,Biochemistry ,Angiodysplasia ,Mice ,03 medical and health sciences ,Conditional gene knockout ,medicine ,Animals ,Molecular Biology ,Mice, Knockout ,Fetus ,Embryogenesis ,Endothelial Cells ,Embryo ,General Medicine ,medicine.disease ,Embryonic stem cell ,Molecular biology ,Cell biology ,030104 developmental biology ,Preclinical imaging - Abstract
Protein arginine methyltransferase 1 (PRMT1) is involved in multiple cellular functions including proliferation and differentiation. Although PRMT1 is expressed in vascular endothelial cells (ECs), which are responsible for angiogenesis during embryonic development, its role has remained elusive. In this study, we generated endothelial-specific prmt1-knockout (Prmt1-ECKO) mice, and found that they died before embryonic day 15. The superficial temporal arteries in these embryos were poorly perfused with blood, and whole-mount 3D imaging revealed dilated and segmentalized luminal structures in Prmt1-ECKO fetuses in comparison with those of controls. Our findings provide evidence that PRMT1 is important for embryonic vascular formation.
- Published
- 2016
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