40 results on '"Perissi, V."'
Search Results
2. Mfn2 deletion in brown adipose tissue protects from insulin resistance and impairs thermogenesis
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Mahdaviani, K, Benador, Iy, Su, S, Gharakhanian, Ra, Stiles, L, Trudeau, Km, Cardamone, M, Enríquez-Zarralanga, V, Ritou, E, Aprahamian, T, Oliveira, Mf, Corkey, Be, Perissi, V, Liesa, M, and Shirihai, Os
- Published
- 2017
3. Clonally expanded mtDNA deletions in human skeletal muscle originate as a proliferative perinuclear niche
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Vincent, A.E., primary, Rosa, H.S., additional, Pabis, K., additional, Lawless, C., additional, Grünewald, A., additional, Chen, C., additional, Rygiel, K.A., additional, Reeve, A.K., additional, Rocha, M.C., additional, Falkous, G., additional, Perissi, V., additional, McWilliams, T.G., additional, Ganley, I.G., additional, White, K., additional, Davey, T., additional, Petrof, B.J., additional, Sayer, A.A., additional, Cooper, C., additional, Taylor, R.W., additional, Turnbull, D.M., additional, and Picard, M., additional
- Published
- 2018
- Full Text
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4. Abstract P2-08-02: Novel tumor suppressor regulating the PI3K/AKT pathway in breast cancer
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Chan, S, primary, Cardamone, MD, additional, and Perissi, V, additional
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- 2018
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5. Molecular determinants of nuclear receptor-corepressor interaction
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Perissi, V., primary, Staszewski, L. M., additional, McInerney, E. M., additional, Kurokawa, R., additional, Krones, A., additional, Rose, D. W., additional, Lambert, M. H., additional, Milburn, M. V., additional, Glass, C. K., additional, and Rosenfeld, M. G., additional
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- 1999
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6. Inhibition of K63 ubiquitination by G-Protein pathway suppressor 2 (GPS2) regulates mitochondria-associated translation.
- Author
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Gao Y, Kwan J, Orofino J, Burrone G, Mitra S, Fan TY, English J, Hekman R, Emili A, Lyons SM, Cardamone MD, and Perissi V
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- Animals, Humans, Mice, Ubiquitin-Conjugating Enzymes metabolism, Ubiquitin-Conjugating Enzymes genetics, Cell Line, Tumor, Mitochondrial Proteins metabolism, Mitochondrial Proteins genetics, Intracellular Signaling Peptides and Proteins, Ubiquitination, Mitochondria metabolism, Protein Biosynthesis
- Abstract
G-Protein Pathway Suppressor 2 (GPS2) is an inhibitor of non-proteolytic K63 ubiquitination mediated by the E2 ubiquitin-conjugating enzyme Ubc13. Previous studies have associated GPS2-mediated restriction of ubiquitination with the regulation of insulin signaling, inflammatory responses and mitochondria-nuclear communication across different tissues and cell types. However, a detailed understanding of the targets of GPS2/Ubc13 activity is lacking. Here, we have dissected the GPS2-regulated K63 ubiquitome in mouse embryonic fibroblasts and human breast cancer cells, unexpectedly finding an enrichment for proteins involved in RNA binding and translation on the outer mitochondrial membrane. Validation of selected targets of GPS2-mediated regulation, including the RNA-binding protein PABPC1 and translation factors RPS1, RACK1 and eIF3M, revealed a mitochondrial-specific strategy for regulating the translation of nuclear-encoded mitochondrial proteins via non-proteolytic ubiquitination. Removal of GPS2-mediated inhibition, either via genetic deletion or stress-induced nuclear translocation, promotes the import-coupled translation of selected mRNAs leading to the increased expression of an adaptive antioxidant program. In light of GPS2 role in nuclear-mitochondria communication, these findings reveal an exquisite regulatory network for modulating mitochondrial gene expression through spatially coordinated transcription and translation., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)
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- 2024
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7. Editorial: Epigenetics and metabolism.
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Mitro N, Verdeguer F, and Perissi V
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- Epigenesis, Genetic, Histones metabolism, Chromatin
- Abstract
Competing Interests: Author FV was employed by the company InSphero AG. 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. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision.
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- 2024
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8. DOT1L stimulates MYC/Mondo transcription factor activity by promoting its degradation cycle on chromatin.
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Sepulveda GP, Gushchanskaia ES, Mora-Martin A, Esse R, Nikorich I, Ceballos A, Kwan J, Blum BC, Dholiya P, Emili A, Perissi V, Cardamone MD, and Grishok A
- Abstract
The proto-oncogene c-MYC is a key representative of the MYC transcription factor network regulating growth and metabolism. MML-1 (Myc- and Mondo-like) is its homolog in C. elegans . The functional and molecular cooperation between c-MYC and H3 lysine 79 methyltransferase DOT1L was demonstrated in several human cancer types, and we have earlier discovered the connection between C. elegans MML-1 and DOT-1.1. Here, we demonstrate the critical role of DOT1L/DOT-1.1 in regulating c-MYC/MML-1 target genes genome-wide by ensuring the removal of "spent" transcription factors from chromatin by the nuclear proteasome. Moreover, we uncover a previously unrecognized proteolytic activity of DOT1L, which may facilitate c-MYC turnover. This new mechanism of c-MYC regulation by DOT1L may lead to the development of new approaches for cancer treatment., Competing Interests: Conflict of interest The authors declare no competing interests.
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- 2024
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9. Multi-Omics Profiling of Hypertrophic Cardiomyopathy Reveals Altered Mechanisms in Mitochondrial Dynamics and Excitation-Contraction Coupling.
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Moore J, Ewoldt J, Venturini G, Pereira AC, Padilha K, Lawton M, Lin W, Goel R, Luptak I, Perissi V, Seidman CE, Seidman J, Chin MT, Chen C, and Emili A
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- Young Adult, Humans, Mitochondrial Dynamics, Multiomics, Proteomics, Myocytes, Cardiac metabolism, Mutation, Cardiomyopathy, Hypertrophic genetics, Induced Pluripotent Stem Cells metabolism
- Abstract
Hypertrophic cardiomyopathy is one of the most common inherited cardiomyopathies and a leading cause of sudden cardiac death in young adults. Despite profound insights into the genetics, there is imperfect correlation between mutation and clinical prognosis, suggesting complex molecular cascades driving pathogenesis. To investigate this, we performed an integrated quantitative multi-omics (proteomic, phosphoproteomic, and metabolomic) analysis to illuminate the early and direct consequences of mutations in myosin heavy chain in engineered human induced pluripotent stem-cell-derived cardiomyocytes relative to late-stage disease using patient myectomies. We captured hundreds of differential features, which map to distinct molecular mechanisms modulating mitochondrial homeostasis at the earliest stages of pathobiology, as well as stage-specific metabolic and excitation-coupling maladaptation. Collectively, this study fills in gaps from previous studies by expanding knowledge of the initial responses to mutations that protect cells against the early stress prior to contractile dysfunction and overt disease.
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- 2023
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10. GPS2-mediated regulation of the adipocyte secretome modulates adipose tissue remodeling at the onset of diet-induced obesity.
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English J, Orofino J, Cederquist CT, Paul I, Li H, Auwerx J, Emili A, Belkina A, Cardamone D, and Perissi V
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- Animals, Mice, Adipocytes metabolism, Obesity metabolism, Adipose Tissue metabolism, Diet, Fibrosis, Intracellular Signaling Peptides and Proteins metabolism, Diabetes Mellitus, Type 2 metabolism
- Abstract
Objective: Dysfunctional, unhealthy expansion of white adipose tissue due to excess dietary intake is a process at the root of obesity and Type 2 Diabetes development. The objective of this study is to contribute to a better understanding of the underlying mechanism(s) regulating the early stages of adipose tissue expansion and adaptation to dietary stress due to an acute, high-fat diet (HFD) challenge, with a focus on the communication between adipocytes and other stromal cells., Methods: We profiled the early response to high-fat diet exposure in wildtype and adipocyte-specific GPS2-KO (GPS2-AKO) mice at the cellular, tissue and organismal level. A multi-pronged approach was employed to disentangle the complex cellular interactions dictating tissue remodeling, via single-cell RNA sequencing and FACS profiling of the stromal fraction, and semi-quantitative proteomics of the adipocyte-derived exosomal cargo after 5 weeks of HFD feeding., Results: Our results indicate that loss of GPS2 in mature adipocytes leads to impaired adaptation to the metabolic stress imposed by HFD feeding. GPS2-AKO mice are significantly more inflamed, insulin resistant, and obese, compared to the WT counterparts. At the cellular level, lack of GPS2 in adipocytes impacts upon other stromal populations, with both the eWAT and scWAT depots exhibiting changes in the immune and non-immune compartments that contribute to an increase in inflammatory and anti-adipogenic cell types. Our studies also revealed that adipocyte to stromal cell communication is facilitated by exosomes, and that transcriptional rewiring of the exosomal cargo is crucial for tissue remodeling. Loss of GPS2 results in increased expression of secreted factors promoting a TGFβ-driven fibrotic microenvironment favoring unhealthy tissue remodeling and expansion., Conclusions: Adipocytes serve as an intercellular signaling hub, communicating with the stromal compartment via paracrine signaling. Our study highlights the importance of proper regulation of the 'secretome' released by energetically stressed adipocytes at the onset of obesity. Altered transcriptional regulation of factors secreted via adipocyte-derived exosomes (AdExos), in the absence of GPS2, contributes to the establishment of an anti-adipogenic, pro-fibrotic adipose tissue environment, and to hastened progression towards a metabolically dysfunctional phenotype., (Copyright © 2023 The Author(s). Published by Elsevier GmbH.. All rights reserved.)
- Published
- 2023
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11. Neuralized-like protein 4 (NEURL4) mediates ADP-ribosylation of mitochondrial proteins.
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Cardamone MD, Gao Y, Kwan J, Hayashi V, Sheeran M, Xu J, English J, Orofino J, Emili A, and Perissi V
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- DNA, Mitochondrial metabolism, HeLa Cells, Homeostasis, Humans, Protein Domains, Ubiquitin-Protein Ligases chemistry, ADP-Ribosylation, Mitochondrial Proteins metabolism, Ubiquitin-Protein Ligases metabolism
- Abstract
ADP-ribosylation is a reversible post-translational modification where an ADP-ribose moiety is covalently attached to target proteins by ADP-ribosyltransferases (ARTs). Although best known for its nuclear roles, ADP-ribosylation is increasingly recognized as a key regulatory strategy across cellular compartments. ADP-ribosylation of mitochondrial proteins has been widely reported, but the exact nature of mitochondrial ART enzymes is debated. We have identified neuralized-like protein 4 (NEURL4) as a mitochondrial ART enzyme and show that most ART activity associated with mitochondria is lost in the absence of NEURL4. The NEURL4-dependent ADP-ribosylome in mitochondrial extracts from HeLa cells includes numerous mitochondrial proteins previously shown to be ADP-ribosylated. In particular, we show that NEURL4 is required for the regulation of mtDNA integrity via poly-ADP-ribosylation of mtLIG3, the rate-limiting enzyme for base excision repair (BER). Collectively, our studies reveal that NEURL4 acts as the main mitochondrial ART enzyme under physiological conditions and provide novel insights in the regulation of mitochondria homeostasis through ADP-ribosylation., (© 2022 Cardamone et al.)
- Published
- 2022
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12. Loss of G-Protein Pathway Suppressor 2 Promotes Tumor Growth Through Activation of AKT Signaling.
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Chan S, Smith E, Gao Y, Kwan J, Blum BC, Tilston-Lunel AM, Turcinovic I, Varelas X, Cardamone MD, Monti S, Emili A, and Perissi V
- Abstract
G Protein Suppressor 2 (GPS2) is a multifunctional protein that exerts important roles in inflammation and metabolism in adipose, liver, and immune cells. GPS2 has recently been identified as a significantly mutated gene in breast cancer and other malignancies and proposed to work as a putative tumor suppressor. However, molecular mechanisms by which GPS2 prevents cancer development and/or progression are largely unknown. Here, we have profiled the phenotypic changes induced by GPS2 depletion in MDA-MB-231 triple negative breast cancer cells and investigated the underlying molecular mechanisms. We found that GPS2-deleted MDA-MB-231 cells exhibited increased proliferative, migratory, and invasive properties in vitro , and conferred greater tumor burden in vivo in an orthotopic xenograft mouse model. Transcriptomic, proteomic and phospho-proteomic profiling of GPS2-deleted MBA-MB-231 revealed a network of altered signals that relate to cell growth and PI3K/AKT signaling. Overlay of GPS2-regulated gene expression with MDA-MB-231 cells modified to express constitutively active AKT showed significant overlap, suggesting that sustained AKT activation is associated with loss of GPS2. Accordingly, we demonstrate that the pro-oncogenic phenotypes associated with GPS2 deletion are rescued by pharmacological inhibition of AKT with MK2206. Collectively, these observations confirm a tumor suppressor role for GPS2 and reveal that loss of GPS2 promotes breast cancer cell proliferation and tumor growth through uncontrolled activation of AKT signaling. Moreover, our study points to GPS2 as a potential biomarker for a subclass of breast cancers that would be responsive to PI3K-class inhibitor drugs., Competing Interests: The 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 © 2021 Chan, Smith, Gao, Kwan, Blum, Tilston-Lunel, Turcinovic, Varelas, Cardamone, Monti, Emili and Perissi.)
- Published
- 2021
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13. Actionable Cytopathogenic Host Responses of Human Alveolar Type 2 Cells to SARS-CoV-2.
- Author
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Hekman RM, Hume AJ, Goel RK, Abo KM, Huang J, Blum BC, Werder RB, Suder EL, Paul I, Phanse S, Youssef A, Alysandratos KD, Padhorny D, Ojha S, Mora-Martin A, Kretov D, Ash PEA, Verma M, Zhao J, Patten JJ, Villacorta-Martin C, Bolzan D, Perea-Resa C, Bullitt E, Hinds A, Tilston-Lunel A, Varelas X, Farhangmehr S, Braunschweig U, Kwan JH, McComb M, Basu A, Saeed M, Perissi V, Burks EJ, Layne MD, Connor JH, Davey R, Cheng JX, Wolozin BL, Blencowe BJ, Wuchty S, Lyons SM, Kozakov D, Cifuentes D, Blower M, Kotton DN, Wilson AA, Mühlberger E, and Emili A
- Published
- 2021
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14. Actionable Cytopathogenic Host Responses of Human Alveolar Type 2 Cells to SARS-CoV-2.
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Hekman RM, Hume AJ, Goel RK, Abo KM, Huang J, Blum BC, Werder RB, Suder EL, Paul I, Phanse S, Youssef A, Alysandratos KD, Padhorny D, Ojha S, Mora-Martin A, Kretov D, Ash PEA, Verma M, Zhao J, Patten JJ, Villacorta-Martin C, Bolzan D, Perea-Resa C, Bullitt E, Hinds A, Tilston-Lunel A, Varelas X, Farhangmehr S, Braunschweig U, Kwan JH, McComb M, Basu A, Saeed M, Perissi V, Burks EJ, Layne MD, Connor JH, Davey R, Cheng JX, Wolozin BL, Blencowe BJ, Wuchty S, Lyons SM, Kozakov D, Cifuentes D, Blower M, Kotton DN, Wilson AA, Mühlberger E, and Emili A
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- Alveolar Epithelial Cells pathology, Alveolar Epithelial Cells virology, Animals, Antiviral Agents, COVID-19 genetics, COVID-19 pathology, Chlorocebus aethiops, Cytopathogenic Effect, Viral, Cytoskeleton, Drug Evaluation, Preclinical, Humans, Induced Pluripotent Stem Cells metabolism, Induced Pluripotent Stem Cells pathology, Induced Pluripotent Stem Cells virology, Phosphoproteins genetics, Protein Transport, Proteome genetics, SARS-CoV-2 genetics, Signal Transduction, Vero Cells, COVID-19 Drug Treatment, Alveolar Epithelial Cells metabolism, COVID-19 metabolism, Phosphoproteins metabolism, Proteome metabolism, SARS-CoV-2 metabolism
- Abstract
Human transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), causative pathogen of the COVID-19 pandemic, exerts a massive health and socioeconomic crisis. The virus infects alveolar epithelial type 2 cells (AT2s), leading to lung injury and impaired gas exchange, but the mechanisms driving infection and pathology are unclear. We performed a quantitative phosphoproteomic survey of induced pluripotent stem cell-derived AT2s (iAT2s) infected with SARS-CoV-2 at air-liquid interface (ALI). Time course analysis revealed rapid remodeling of diverse host systems, including signaling, RNA processing, translation, metabolism, nuclear integrity, protein trafficking, and cytoskeletal-microtubule organization, leading to cell cycle arrest, genotoxic stress, and innate immunity. Comparison to analogous data from transformed cell lines revealed respiratory-specific processes hijacked by SARS-CoV-2, highlighting potential novel therapeutic avenues that were validated by a high hit rate in a targeted small molecule screen in our iAT2 ALI system., Competing Interests: Declaration of Interests B.L.W. declares a position as CSO of Aquinnah Pharmaceuticals. A.E. and D.N.K. declare industry funding from Johnson & Johnson, Merck, and Novartis., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2020
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15. Decoding the rosetta stone of mitonuclear communication.
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English J, Son JM, Cardamone MD, Lee C, and Perissi V
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- Animals, Cell Nucleus genetics, Gene Expression Regulation, Humans, Mitochondria genetics, Mitochondrial Proteins genetics, Nuclear Proteins genetics, Signal Transduction, Cell Communication, Cell Nucleus metabolism, Mitochondria metabolism, Mitochondrial Proteins metabolism, Nuclear Proteins metabolism
- Abstract
Cellular homeostasis in eukaryotic cells requires synchronized coordination of multiple organelles. A key role in this stage is played by mitochondria, which have recently emerged as highly interconnected and multifunctional hubs that process and coordinate diverse cellular functions. Beyond producing ATP, mitochondria generate key metabolites and are central to apoptotic and metabolic signaling pathways. Because most mitochondrial proteins are encoded in the nuclear genome, the biogenesis of new mitochondria and the maintenance of mitochondrial functions and flexibility critically depend upon effective mitonuclear communication. This review addresses the complex network of signaling molecules and pathways allowing mitochondria-nuclear communication and coordinated regulation of their independent but interconnected genomes, and discusses the extent to which dynamic communication between the two organelles has evolved for mutual benefit and for the overall maintenance of cellular and organismal fitness., (Copyright © 2020 Elsevier Ltd. All rights reserved.)
- Published
- 2020
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16. The Adipocyte Acquires a Fibroblast-Like Transcriptional Signature in Response to a High Fat Diet.
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Jones JEC, Rabhi N, Orofino J, Gamini R, Perissi V, Vernochet C, and Farmer SR
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- Animals, Extracellular Matrix Proteins genetics, Extracellular Matrix Proteins metabolism, Fibroblasts metabolism, Intra-Abdominal Fat cytology, Male, Mice, Mice, Inbred C57BL, Adipocytes, White metabolism, Diet, High-Fat adverse effects, Intra-Abdominal Fat metabolism, Transcriptome
- Abstract
Visceral white adipose tissue (vWAT) expands and undergoes extensive remodeling during diet-induced obesity. Much is known about the contribution of various stromal vascular cells to the remodeling process, but less is known of the changes that occur within the adipocyte as it becomes progressively dysfunctional. Here, we performed a transcriptome analysis of isolated vWAT adipocytes to assess global pathway changes occurring in response to a chronic high fat diet (HFD). The data demonstrate that the adipocyte responds to the HFD by adopting a fibroblast-like phenotype, characterized by enhanced expression of ECM, focal adhesion and cytoskeletal genes and suppression of many adipocyte programs most notably those associated with mitochondria. This study reveals that during obesity the adipocyte progressively becomes metabolically dysfunctional due to its acquisition of fibrogenic functions. We propose that mechano-responsive transcription factors such as MRTFA and SRF contribute to both upregulation of morphological genes as well as suppression of mitochondrial programs.
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- 2020
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17. Opposing action of NCoR1 and PGC-1α in mitochondrial redox homeostasis.
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Lima TI, Guimarães DSPSF, Oliveira AG, Araujo H, Sponton CHG, Souza-Pinto NC, Saito Â, Figueira ACM, Palameta S, Bajgelman MC, Calixto A, Pinto S, Mori MA, Orofino J, Perissi V, Mottis A, Auwerx J, and Silveira LR
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- Animals, Antioxidants metabolism, Caenorhabditis elegans, Cell Survival, Green Fluorescent Proteins metabolism, Homeostasis, Lipids chemistry, Mice, Muscle, Skeletal metabolism, Palmitates pharmacology, Propidium pharmacology, RNA, Small Interfering metabolism, Reactive Oxygen Species metabolism, Superoxide Dismutase metabolism, Trans-Activators metabolism, Transcription, Genetic, Gene Expression Regulation, Mitochondria metabolism, Nuclear Receptor Co-Repressor 1 metabolism, Oxidation-Reduction drug effects, Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha metabolism
- Abstract
The ability to respond to fluctuations of reactive oxygen species (ROS) within the cell is a central aspect of mammalian physiology. This dynamic process depends on the coordinated action of transcriptional factors to promote the expression of genes encoding for antioxidant enzymes. Here, we demonstrate that the transcriptional coregulators, PGC-1α and NCoR1, are essential mediators of mitochondrial redox homeostasis in skeletal muscle cells. Our findings reveal an antagonistic role of these coregulators in modulating mitochondrial antioxidant induction through Sod2 transcriptional control. Importantly, the activation of this mechanism by either PGC-1α overexpression or NCoR1 knockdown attenuates mitochondrial ROS levels and prevents cell death caused by lipid overload in skeletal muscle cells. The opposing actions of coactivators and corepressors, therefore, exert a commanding role over cellular antioxidant capacity., (Copyright © 2019 Elsevier Inc. All rights reserved.)
- Published
- 2019
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18. Chromatin Immunoprecipitation of Murine Brown Adipose Tissue.
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Cardamone MD, Orofino J, Labadorf A, and Perissi V
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- Adipose Tissue, Brown chemistry, Animals, Cell Nucleus metabolism, DNA genetics, DNA metabolism, Epigenomics methods, Histone Code physiology, Mice, Protein Processing, Post-Translational physiology, Sequence Analysis, DNA methods, Transcription Factors genetics, Transcription Factors metabolism, Adipose Tissue, Brown physiology, Chromatin Immunoprecipitation methods, Protein Array Analysis methods
- Abstract
Most cellular processes are regulated by transcriptional modulation of specific gene programs. Such modulation is achieved through the combined actions of a wide range of transcription factors (TFs) and cofactors mediating transcriptional activation or repression via changes in chromatin structure. Chromatin immunoprecipitation (ChIP) is a useful molecular biology approach for mapping histone modifications and profiling transcription factors/cofactors binding to DNA, thus providing a snapshot of the dynamic nuclear changes occurring during different biological processes. To study transcriptional regulation in adipose tissue, samples derived from in vitro cell cultures of immortalized or primary cell lines are often favored in ChIP assays because of the abundance of starting material and reduced biological variability. However, these models represent a limited snapshot of the actual chromatin state in living organisms. Thus, there is a critical need for optimized protocols to perform ChIP on adipose tissue samples derived from animal models. Here we describe a protocol for efficient ChIP-seq of both histone modifications and non-histone proteins in brown adipose tissue (BAT) isolated from a mouse. The protocol is optimized for investigating genome-wide localization of proteins of interest and epigenetic markers in the BAT, which is a morphologically and physiologically distinct tissue amongst fat depots.
- Published
- 2018
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19. Subcellular origin of mitochondrial DNA deletions in human skeletal muscle.
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Vincent AE, Rosa HS, Pabis K, Lawless C, Chen C, Grünewald A, Rygiel KA, Rocha MC, Reeve AK, Falkous G, Perissi V, White K, Davey T, Petrof BJ, Sayer AA, Cooper C, Deehan D, Taylor RW, Turnbull DM, and Picard M
- Subjects
- Aging pathology, Humans, Muscle Fibers, Skeletal pathology, Muscle Fibers, Skeletal ultrastructure, Muscle, Skeletal pathology, Subcellular Fractions pathology, Subcellular Fractions ultrastructure, Aging genetics, DNA, Mitochondrial genetics, DNA, Mitochondrial ultrastructure, Gene Deletion, Muscle, Skeletal physiology, Muscle, Skeletal ultrastructure
- Abstract
Objective: In patients with mitochondrial DNA (mtDNA) maintenance disorders and with aging, mtDNA deletions sporadically form and clonally expand within individual muscle fibers, causing respiratory chain deficiency. This study aimed to identify the sub-cellular origin and potential mechanisms underlying this process., Methods: Serial skeletal muscle cryosections from patients with multiple mtDNA deletions were subjected to subcellular immunofluorescent, histochemical, and genetic analysis., Results: We report respiratory chain-deficient perinuclear foci containing mtDNA deletions, which show local elevations of both mitochondrial mass and mtDNA copy number. These subcellular foci of respiratory chain deficiency are associated with a local increase in mitochondrial biogenesis and unfolded protein response signaling pathways. We also find that the commonly reported segmental pattern of mitochondrial deficiency is consistent with the three-dimensional organization of the human skeletal muscle mitochondrial network., Interpretation: We propose that mtDNA deletions first exceed the biochemical threshold causing biochemical deficiency in focal regions adjacent to the myonuclei, and induce mitochondrial biogenesis before spreading across the muscle fiber. These subcellular resolution data provide new insights into the possible origin of mitochondrial respiratory chain deficiency in mitochondrial myopathy. Ann Neurol 2018;84:289-301., (© 2018 The Authors. Annals of Neurology published by Wiley Periodicals, Inc. on behalf of American Neurological Association.)
- Published
- 2018
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20. Mitochondrial Retrograde Signaling in Mammals Is Mediated by the Transcriptional Cofactor GPS2 via Direct Mitochondria-to-Nucleus Translocation.
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Cardamone MD, Tanasa B, Cederquist CT, Huang J, Mahdaviani K, Li W, Rosenfeld MG, Liesa M, and Perissi V
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- 3T3-L1 Cells, Active Transport, Cell Nucleus physiology, Animals, Cell Nucleus genetics, HeLa Cells, Histones genetics, Histones metabolism, Humans, Intracellular Signaling Peptides and Proteins genetics, Methylation, Mice, Mitochondria genetics, Promoter Regions, Genetic physiology, Transcriptional Activation physiology, Cell Nucleus metabolism, Intracellular Signaling Peptides and Proteins metabolism, Mitochondria metabolism, Organelle Biogenesis, Signal Transduction physiology
- Abstract
As most of the mitochondrial proteome is encoded in the nucleus, mitochondrial functions critically depend on nuclear gene expression and bidirectional mito-nuclear communication. However, mitochondria-to-nucleus communication pathways in mammals are incompletely understood. Here, we identify G-Protein Pathway Suppressor 2 (GPS2) as a mediator of mitochondrial retrograde signaling and a transcriptional activator of nuclear-encoded mitochondrial genes. GPS2-regulated translocation from mitochondria to nucleus is essential for the transcriptional activation of a nuclear stress response to mitochondrial depolarization and for supporting basal mitochondrial biogenesis in differentiating adipocytes and brown adipose tissue (BAT) from mice. In the nucleus, GPS2 recruitment to target gene promoters regulates histone H3K9 demethylation and RNA POL2 activation through inhibition of Ubc13-mediated ubiquitination. These findings, together, reveal an additional layer of regulation of mitochondrial gene transcription, uncover a direct mitochondria-nuclear communication pathway, and indicate that GPS2 retrograde signaling is a key component of the mitochondrial stress response in mammals., (Copyright © 2018 Elsevier Inc. All rights reserved.)
- Published
- 2018
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21. Mfn2 deletion in brown adipose tissue protects from insulin resistance and impairs thermogenesis.
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Mahdaviani K, Benador IY, Su S, Gharakhanian RA, Stiles L, Trudeau KM, Cardamone M, Enríquez-Zarralanga V, Ritou E, Aprahamian T, Oliveira MF, Corkey BE, Perissi V, Liesa M, and Shirihai OS
- Subjects
- Animals, Diet, High-Fat, Energy Metabolism, Female, Glycolysis, Male, Mice, Mitochondria metabolism, Mitochondrial Proteins metabolism, Obesity, Adipose Tissue, Brown metabolism, GTP Phosphohydrolases deficiency, GTP Phosphohydrolases genetics, Insulin Resistance, Thermogenesis genetics
- Abstract
BAT-controlled thermogenic activity is thought to be required for its capacity to prevent the development of insulin resistance. This hypothesis predicts that mediators of thermogenesis may help prevent diet-induced insulin resistance. We report that the mitochondrial fusion protein Mitofusin 2 (Mfn2) in BAT is essential for cold-stimulated thermogenesis, but promotes insulin resistance in obese mice. Mfn2 deletion in mice through Ucp1-cre (BAT-Mfn2-KO) causes BAT lipohypertrophy and cold intolerance. Surprisingly however, deletion of Mfn2 in mice fed a high fat diet (HFD) results in improved insulin sensitivity and resistance to obesity, while impaired cold-stimulated thermogenesis is maintained. Improvement in insulin sensitivity is associated with a gender-specific remodeling of BAT mitochondrial function. In females, BAT mitochondria increase their efficiency for ATP-synthesizing fat oxidation, whereas in BAT from males, complex I-driven respiration is decreased and glycolytic capacity is increased. Thus, BAT adaptation to obesity is regulated by Mfn2 and with BAT-Mfn2 absent, BAT contribution to prevention of insulin resistance is independent and inversely correlated to whole-body cold-stimulated thermogenesis., (© 2017 The Authors. Published under the terms of the CC BY 4.0 license.)
- Published
- 2017
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22. Inhibition of Ubc13-mediated Ubiquitination by GPS2 Regulates Multiple Stages of B Cell Development.
- Author
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Lentucci C, Belkina AC, Cederquist CT, Chan M, Johnson HE, Prasad S, Lopacinski A, Nikolajczyk BS, Monti S, Snyder-Cappione J, Tanasa B, Cardamone MD, and Perissi V
- Subjects
- Animals, Bone Marrow Cells cytology, Cell Differentiation, Gene Expression Profiling, Mice, Mice, Inbred C57BL, Mice, Knockout, Receptors, Antigen, B-Cell metabolism, Signal Transduction, Ubiquitination, B-Lymphocytes cytology, Intracellular Signaling Peptides and Proteins metabolism, Ubiquitin-Conjugating Enzymes antagonists & inhibitors
- Abstract
Non-proteolytic ubiquitin signaling mediated by Lys
63 ubiquitin chains plays a critical role in multiple pathways that are key to the development and activation of immune cells. Our previous work indicates that GPS2 (G-protein Pathway Suppressor 2) is a multifunctional protein regulating TNFα signaling and lipid metabolism in the adipose tissue through modulation of Lys63 ubiquitination events. However, the full extent of GPS2-mediated regulation of ubiquitination and the underlying molecular mechanisms are unknown. Here, we report that GPS2 is required for restricting the activation of TLR and BCR signaling pathways and the AKT/FOXO1 pathway in immune cells based on direct inhibition of Ubc13 enzymatic activity. Relevance of this regulatory strategy is confirmed in vivo by B cell-targeted deletion of GPS2, resulting in developmental defects at multiple stages of B cell differentiation. Together, these findings reveal that GPS2 genomic and non-genomic functions are critical for the development and cellular homeostasis of B cells., (© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.)- Published
- 2017
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23. Systemic insulin sensitivity is regulated by GPS2 inhibition of AKT ubiquitination and activation in adipose tissue.
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Cederquist CT, Lentucci C, Martinez-Calejman C, Hayashi V, Orofino J, Guertin D, Fried SK, Lee MJ, Cardamone MD, and Perissi V
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- 3T3 Cells, Adipocytes metabolism, Animals, Diabetes Mellitus, Type 2 metabolism, Inflammation genetics, Insulin genetics, Insulin physiology, Insulin Resistance genetics, Intracellular Signaling Peptides and Proteins metabolism, Mice, Obesity genetics, Proto-Oncogene Proteins c-akt metabolism, Signal Transduction, Ubiquitin-Conjugating Enzymes antagonists & inhibitors, Ubiquitin-Conjugating Enzymes metabolism, Ubiquitination, Adipose Tissue metabolism, Insulin metabolism, Intracellular Signaling Peptides and Proteins physiology
- Abstract
Objective: Insulin signaling plays a unique role in the regulation of energy homeostasis and the impairment of insulin action is associated with altered lipid metabolism, obesity, and Type 2 Diabetes. The main aim of this study was to provide further insight into the regulatory mechanisms governing the insulin signaling pathway by investigating the role of non-proteolytic ubiquitination in insulin-mediated activation of AKT., Methods: The molecular mechanism of AKT regulation through ubiquitination is first dissected in vitro in 3T3-L1 preadipocytes and then validated in vivo using mice with adipo-specific deletion of GPS2, an endogenous inhibitor of Ubc13 activity (GPS2-AKO mice)., Results: Our results indicate that K63 ubiquitination is a critical component of AKT activation in the insulin signaling pathway and that counter-regulation of this step is provided by GPS2 preventing AKT ubiquitination through inhibition of Ubc13 enzymatic activity. Removal of this negative checkpoint, through GPS2 downregulation or genetic deletion, results in sustained activation of insulin signaling both in vitro and in vivo . As a result, the balance between lipid accumulation and utilization is shifted toward storage in the adipose tissue and GPS2-AKO mice become obese under normal laboratory chow diet. However, the adipose tissue of GPS2-AKO mice is not inflamed, the levels of circulating adiponectin are elevated, and systemic insulin sensitivity is overall improved., Conclusions: Our findings characterize a novel layer of regulation of the insulin signaling pathway based on non-proteolytic ubiquitination of AKT and define GPS2 as a previously unrecognized component of the insulin signaling cascade. In accordance with this role, we have shown that GPS2 presence in adipocytes modulates systemic metabolism by restricting the activation of insulin signaling during the fasted state, whereas in absence of GPS2, the adipose tissue is more efficient at lipid storage, and obesity becomes uncoupled from inflammation and insulin resistance.
- Published
- 2016
- Full Text
- View/download PDF
24. Exchange Factor TBL1 and Arginine Methyltransferase PRMT6 Cooperate in Protecting G Protein Pathway Suppressor 2 (GPS2) from Proteasomal Degradation.
- Author
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Huang J, Cardamone MD, Johnson HE, Neault M, Chan M, Floyd ZE, Mallette FA, and Perissi V
- Subjects
- Active Transport, Cell Nucleus, HEK293 Cells, HeLa Cells, Humans, Intracellular Signaling Peptides and Proteins chemistry, Methylation, Nuclear Localization Signals, Protein Stability, Protein Structure, Tertiary, Proteolysis, Ubiquitination, Intracellular Signaling Peptides and Proteins metabolism, Nuclear Proteins physiology, Proteasome Endopeptidase Complex metabolism, Protein-Arginine N-Methyltransferases physiology, Transducin physiology
- Abstract
G protein pathway suppressor 2 (GPS2) is a multifunctional protein involved in the regulation of a number of metabolic organs. First identified as part of the NCoR-SMRT corepressor complex, GPS2 is known to play an important role in the nucleus in the regulation of gene transcription and meiotic recombination. In addition, we recently reported a non-transcriptional role of GPS2 as an inhibitor of the proinflammatory TNFα pathway in the cytosol. Although this suggests that the control of GPS2 localization may be an important determinant of its molecular functions, a clear understanding of GPS2 differential targeting to specific cellular locations is still lacking. Here we show that a fine balance between protein stabilization and degradation tightly regulates GPS2 nuclear function. Our findings indicate that GPS2 is degraded upon polyubiquitination by the E3 ubiquitin ligase Siah2. Unexpectedly, interaction with the exchange factor TBL1 is required to protect GPS2 from degradation, with methylation of GPS2 by arginine methyltransferase PRMT6 regulating the interaction with TBL1 and inhibiting proteasome-dependent degradation. Overall, our findings indicate that regulation of GPS2 by posttranslational modifications provides an effective strategy for modulating its molecular function within the nuclear compartment., (© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2015
- Full Text
- View/download PDF
25. GPS2/KDM4A pioneering activity regulates promoter-specific recruitment of PPARγ.
- Author
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Cardamone MD, Tanasa B, Chan M, Cederquist CT, Andricovich J, Rosenfeld MG, and Perissi V
- Subjects
- Adipocytes metabolism, Animals, Chromatin Assembly and Disassembly, Lipase genetics, Lipase metabolism, Lipolysis genetics, Mice, Sterol Esterase genetics, Sterol Esterase metabolism, Ubiquitin-Protein Ligases metabolism, Histone Demethylases metabolism, Intracellular Signaling Peptides and Proteins metabolism, PPAR gamma metabolism, Promoter Regions, Genetic
- Abstract
Timely and selective recruitment of transcription factors to their appropriate DNA-binding sites represents a critical step in regulating gene activation; however, the regulatory strategies underlying each factor's effective recruitment to specific promoter and/or enhancer regions are not fully understood. Here, we identify an unexpected regulatory mechanism by which promoter-specific binding, and therefore function, of peroxisome proliferator-activator receptor γ (PPARγ) in adipocytes requires G protein suppressor 2 (GPS2) to prime the local chromatin environment via inhibition of the ubiquitin ligase RNF8 and stabilization of the H3K9 histone demethylase KDM4A/JMJD2. Integration of genome-wide profiling data indicates that the pioneering activity of GPS2/KDM4A is required for PPARγ-mediated regulation of a specific transcriptional program, including the lipolytic enzymes adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL). Hence, our findings reveal that GPS2 exerts a biologically important function in adipose tissue lipid mobilization by directly regulating ubiquitin signaling and indirectly modulating chromatin remodeling to prime selected genes for activation., (Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2014
- Full Text
- View/download PDF
26. The co-repressor SMRT delays DNA damage-induced caspase activation by repressing pro-apoptotic genes and modulating the dynamics of checkpoint kinase 2 activation.
- Author
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Scafoglio C, Smolka M, Zhou H, Perissi V, and Rosenfeld MG
- Subjects
- Apoptosis genetics, Cell Line, Tumor, Chromatin Immunoprecipitation, Enzyme Activation physiology, Fluorescent Antibody Technique, Humans, Microarray Analysis, Protein Phosphatase 2C, Reverse Transcriptase Polymerase Chain Reaction, Apoptosis physiology, Checkpoint Kinase 2 metabolism, DNA Damage physiology, Gene Expression Regulation physiology, Nuclear Receptor Co-Repressor 2 metabolism, Phosphoprotein Phosphatases metabolism
- Abstract
Checkpoint kinase 2 (Chk2) is a major regulator of DNA damage response and can induce alternative cellular responses: cell cycle arrest and DNA repair or programmed cell death. Here, we report the identification of a new role of Chk2 in transcriptional regulation that also contributes to modulating the balance between survival and apoptosis following DNA damage. We found that Chk2 interacts with members of the NCoR/SMRT transcriptional co-regulator complexes and serves as a functional component of the repressor complex, being required for recruitment of SMRT on the promoter of pro-apoptotic genes upon DNA damage. Thus, the co-repressor SMRT exerts a critical protective action against genotoxic stress-induced caspase activation, repressing a functionally important cohort of pro-apoptotic genes. Amongst them, SMRT is responsible for basal repression of Wip1, a phosphatase that de-phosphorylates and inactivates Chk2, thus affecting a feedback loop responsible for licensing the correct timing of Chk2 activation and the proper execution of the DNA repair process.
- Published
- 2013
- Full Text
- View/download PDF
27. A protective strategy against hyperinflammatory responses requiring the nontranscriptional actions of GPS2.
- Author
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Cardamone MD, Krones A, Tanasa B, Taylor H, Ricci L, Ohgi KA, Glass CK, Rosenfeld MG, and Perissi V
- Subjects
- Adipose Tissue immunology, Animals, Cell Line, GTPase-Activating Proteins genetics, GTPase-Activating Proteins immunology, GTPase-Activating Proteins metabolism, Inflammation genetics, Inflammation immunology, Inflammation metabolism, Insulin genetics, Insulin immunology, Insulin metabolism, Intracellular Signaling Peptides and Proteins genetics, Intracellular Signaling Peptides and Proteins immunology, MAP Kinase Kinase 4 genetics, MAP Kinase Kinase 4 immunology, MAP Kinase Kinase 4 metabolism, Macrophages immunology, Mice, Mice, Transgenic, Resistin genetics, Resistin immunology, Resistin metabolism, Signal Transduction genetics, Signal Transduction immunology, Tumor Necrosis Factor-alpha genetics, Tumor Necrosis Factor-alpha immunology, Tumor Necrosis Factor-alpha metabolism, Ubiquitin-Conjugating Enzymes genetics, Ubiquitin-Conjugating Enzymes immunology, Ubiquitin-Conjugating Enzymes metabolism, Ubiquitination genetics, Ubiquitination immunology, Adipose Tissue metabolism, Homeostasis, Intracellular Signaling Peptides and Proteins metabolism, Macrophages metabolism
- Abstract
The association between hyperinflammatory states and numerous diseases is widely recognized, but our understanding of the molecular strategies that have evolved to prevent uncontrolled activation of inflammatory responses remains incomplete. Here, we report a critical, nontranscriptional role of GPS2 as a guardian against hyperstimulation of the TNF-α-induced gene program. GPS2 cytoplasmic actions are required to specifically modulate RIP1 ubiquitylation and JNK activation by inhibiting TRAF2/Ubc13 enzymatic activity. In vivo relevance of GPS2 anti-inflammatory role is confirmed by inhibition of TNF-α target genes in macrophages and by improved insulin signaling in the adipose tissue of aP2-GPS2 transgenic mice. As the nontranscriptional role is complemented by GPS2 functioning as positive and negative cofactor for nuclear receptors, in vivo overexpression also results in elevated circulating level of Resistin and development of hepatic steatosis. Together, these studies define GPS2 as a molecular guardian required for precise control of inflammatory responses involved in immunity and homeostasis., (Copyright © 2012 Elsevier Inc. All rights reserved.)
- Published
- 2012
- Full Text
- View/download PDF
28. Deconstructing repression: evolving models of co-repressor action.
- Author
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Perissi V, Jepsen K, Glass CK, and Rosenfeld MG
- Subjects
- Animals, Binding Sites, Co-Repressor Proteins genetics, DNA chemistry, Gene Expression, Humans, Models, Biological, Repressor Proteins genetics, Repressor Proteins metabolism, Transcription Factors genetics, Co-Repressor Proteins metabolism, Models, Genetic
- Abstract
A crucial aspect of development, homeostasis and prevention of disease is the strict maintenance of patterns of gene repression. Gene repression is largely achieved by the combinatorial action of various enzymatic complexes - known as co-repressor complexes - that are recruited to DNA by transcription factors and often act through enzymatic modification of histone protein tails. Our understanding of how co-repressors act has begun to change over recent years owing to the increased availability of genome-scale data. Here, we consider specific strategies that underlie repression events - for example, those mediated by the nuclear receptor co-repressor (NCoR, also known as NCOR1) and silencing mediator of retinoic acid and thyroid hormone receptor (SMRT, also known as NCOR2) co-repressor complexes - and discuss emerging themes in gene repression.
- Published
- 2010
- Full Text
- View/download PDF
29. Transcriptional integration of TLR2 and TLR4 signaling at the NCoR derepression checkpoint.
- Author
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Huang W, Ghisletti S, Perissi V, Rosenfeld MG, and Glass CK
- Subjects
- Animals, Calcium-Calmodulin-Dependent Protein Kinase Type 2 genetics, Calcium-Calmodulin-Dependent Protein Kinase Type 2 metabolism, Cell Line, Cells, Cultured, Chromatin Immunoprecipitation, DNA-Binding Proteins genetics, DNA-Binding Proteins metabolism, Immunoblotting, Lipopeptides pharmacology, Lipopolysaccharides pharmacology, Liver X Receptors, Mice, Mice, Inbred C57BL, Mutation, Nuclear Proteins genetics, Nuclear Receptor Co-Repressor 1, Nuclear Receptor Co-Repressor 2, Orphan Nuclear Receptors, Phosphorylation drug effects, RNA Interference, Receptors, Cytoplasmic and Nuclear metabolism, Repressor Proteins genetics, Reverse Transcriptase Polymerase Chain Reaction, Tetradecanoylphorbol Acetate pharmacology, Toll-Like Receptor 2 genetics, Toll-Like Receptor 4 genetics, Transcription Factor RelA genetics, Transcription Factor RelA metabolism, Transcription, Genetic drug effects, Nuclear Proteins metabolism, Repressor Proteins metabolism, Signal Transduction, Toll-Like Receptor 2 metabolism, Toll-Like Receptor 4 metabolism
- Abstract
Activation of toll-like receptors (TLRs) leads to derepression and subsequent activation of inflammatory response genes that play essential roles in innate and acquired immunity. Derepression requires signal-dependent turnover of the nuclear receptor corepressor NCoR from target promoters, but the mechanisms remain poorly understood. Here, we report that TLR4 uses NFkappaB to deliver IKKepsilon to target promoters that contain "integrated circuits" of kappaB and AP-1 sites, resulting in local phosphorylation of c-Jun and subsequent NCoR clearance. In contrast, TLR2 signaling leads to rapid activation of CaMKII and phosphorylation of the TBLR1 component of NCoR complexes, bypassing the requirement for c-Jun phosphorylation and enabling NCoR clearance from promoters lacking integrated kappaB elements. Intriguingly, the IKKvarepsilon-dependent clearance pathway is sensitive to transrepression by liver X receptors, while the CaMKII-dependent pathway is not. These findings reveal mechanisms for integration of TLR, calcium, and nuclear receptor signaling pathways that underlie pathogen-specific responses and disease-specific programs of inflammation.
- Published
- 2009
- Full Text
- View/download PDF
30. TBL1 and TBLR1 phosphorylation on regulated gene promoters overcomes dual CtBP and NCoR/SMRT transcriptional repression checkpoints.
- Author
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Perissi V, Scafoglio C, Zhang J, Ohgi KA, Rose DW, Glass CK, and Rosenfeld MG
- Subjects
- Animals, Breast Neoplasms, Cell Line, Cell Line, Tumor, Female, Gene Expression Regulation, Neoplastic, Genes, Reporter, Humans, Nuclear Receptor Co-Repressor 2, Proteasome Endopeptidase Complex metabolism, Repressor Proteins genetics, Transcriptional Activation, Ubiquitin metabolism, Alcohol Oxidoreductases genetics, DNA-Binding Proteins genetics, Nuclear Proteins metabolism, Promoter Regions, Genetic, Receptors, Cytoplasmic and Nuclear metabolism, Repressor Proteins metabolism, Transcription, Genetic, Transducin metabolism
- Abstract
A key strategy to achieve regulated gene expression in higher eukaryotes is to prevent illegitimate signal-independent activation by imposing robust control on the dismissal of corepressors. Here, we report that many signaling pathways, including Notch, NF-kappaB, and nuclear receptor ligands, are subjected to a dual-repression "checkpoint" based on distinct corepressor complexes. Gene activation requires the release of both CtBP1/2- and NCoR/SMRT-dependent repression, through the coordinate action of two highly related exchange factors, the transducer beta-like proteins TBL1 and TBLR1, that license ubiquitylation and degradation of CtBP1/2 and NCoR/SMRT, respectively. Intriguingly, their function and differential specificity reside in only five specific Ser/Thr phosphorylation site differences, regulated by direct phosphorylation at the level of the promoter, as exemplified by the role of PKCdelta in TBLR1-dependent dismissal of NCoR. Thus, our data reveal a strategy of dual-factor repression checkpoints, in which dedicated exchange factors serve as sensors for signal-specific dismissal of distinct corepressors, with specificity imposed by upstream signaling pathways.
- Published
- 2008
- Full Text
- View/download PDF
31. Anti-inflammatory and antidiabetic roles of PPARgamma.
- Author
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Pascual G, Sullivan AL, Ogawa S, Gamliel A, Perissi V, Rosenfeld MG, and Glass CK
- Subjects
- Animals, Diabetes Mellitus, Type 2 drug therapy, Genome, Humans, Inflammation drug therapy, Ligands, Macrophages metabolism, Models, Biological, Nuclear Receptor Coactivator 2 metabolism, Anti-Inflammatory Agents metabolism, Hypoglycemic Agents metabolism, PPAR gamma metabolism
- Abstract
The peroxisome proliferator-activated receptor gamma (PPARgamma) regulates adipocyte differentiation and glucose homeostasis and is the molecular target of thiazolidinediones (TZDs) that act as insulin-sensitizers in patients with type 2 diabetes. PPARgamma is also expressed in macrophages and negatively regulates the programme of macrophage activation by repressing a subset of AP1 and NF-kappaB-dependent genes. Recent genetic, molecular and biochemical studies support the idea that PPARgamma inhibits inflammatory gene expression in activated macrophages by a NCoR/sumoylation-dependent pathway. Sumoylation of PPARgamma targets it to NCoR corepressor complexes that are bound to inflammatory response gene promoters and prevents their signal-dependent clearance that is normally a prerequisite for transcriptional activation. As a consequence, genes remain in a repressed state. Because the ligand-induced allosteric changes that promote entry of PPARgamma into this transrepression pathway are distinct from those that mediate interactions with conventional coactivators, these findings may facilitate the development of novel PPARgamma ligands that retain antidiabetic activities but have reduced side effects.
- Published
- 2007
- Full Text
- View/download PDF
32. A topoisomerase IIbeta-mediated dsDNA break required for regulated transcription.
- Author
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Ju BG, Lunyak VV, Perissi V, Garcia-Bassets I, Rose DW, Glass CK, and Rosenfeld MG
- Subjects
- Cell Line, Tumor, Chromatin metabolism, Chromatin Immunoprecipitation, DNA Damage, DNA Repair, DNA-Binding Proteins antagonists & inhibitors, Enzyme Inhibitors pharmacology, Estradiol pharmacology, Estrogen Receptor alpha metabolism, Histones metabolism, Humans, Membrane Proteins genetics, Nucleosomes metabolism, Poly (ADP-Ribose) Polymerase-1, Poly(ADP-ribose) Polymerases metabolism, Presenilin-2, Promoter Regions, Genetic, Response Elements, Thiobarbiturates pharmacology, Topoisomerase II Inhibitors, Transcription Factors metabolism, Transfection, DNA metabolism, DNA Topoisomerases, Type II metabolism, DNA-Binding Proteins metabolism, Transcription, Genetic, Transcriptional Activation
- Abstract
Multiple enzymatic activities are required for transcriptional initiation. The enzyme DNA topoisomerase II associates with gene promoter regions and can generate breaks in double-stranded DNA (dsDNA). Therefore, it is of interest to know whether this enzyme is critical for regulated gene activation. We report that the signal-dependent activation of gene transcription by nuclear receptors and other classes of DNA binding transcription factors, including activating protein 1, requires DNA topoisomerase IIbeta-dependent, transient, site-specific dsDNA break formation. Subsequent to the break, poly(adenosine diphosphate-ribose) polymerase-1 enzymatic activity is induced, which is required for a nucleosome-specific histone H1-high-mobility group B exchange event and for local changes of chromatin architecture. Our data mechanistically link DNA topoisomerase IIbeta-dependent dsDNA breaks and the components of the DNA damage and repair machinery in regulated gene transcription.
- Published
- 2006
- Full Text
- View/download PDF
33. A SUMOylation-dependent pathway mediates transrepression of inflammatory response genes by PPAR-gamma.
- Author
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Pascual G, Fong AL, Ogawa S, Gamliel A, Li AC, Perissi V, Rose DW, Willson TM, Rosenfeld MG, and Glass CK
- Subjects
- Animals, Cells, Cultured, Histone Deacetylases metabolism, Ligands, Lipopolysaccharides pharmacology, Macrophages metabolism, Mice, Multiprotein Complexes metabolism, NF-kappa B metabolism, Nitric Oxide Synthase genetics, Nitric Oxide Synthase Type II, Nuclear Proteins metabolism, Nuclear Receptor Co-Repressor 1, Protein Binding drug effects, Protein Inhibitors of Activated STAT, Proteins metabolism, Down-Regulation drug effects, Inflammation genetics, PPAR gamma metabolism, Repressor Proteins metabolism, SUMO-1 Protein metabolism
- Abstract
Peroxisome proliferator-activated receptor-gamma (PPAR-gamma) has essential roles in adipogenesis and glucose homeostasis, and is a molecular target of insulin-sensitizing drugs. Although the ability of PPAR-gamma agonists to antagonize inflammatory responses by transrepression of nuclear factor kappa B (NF-kappaB) target genes is linked to antidiabetic and antiatherogenic actions, the mechanisms remain poorly understood. Here we report the identification of a molecular pathway by which PPAR-gamma represses the transcriptional activation of inflammatory response genes in mouse macrophages. The initial step of this pathway involves ligand-dependent SUMOylation of the PPAR-gamma ligand-binding domain, which targets PPAR-gamma to nuclear receptor corepressor (NCoR)-histone deacetylase-3 (HDAC3) complexes on inflammatory gene promoters. This in turn prevents recruitment of the ubiquitylation/19S proteosome machinery that normally mediates the signal-dependent removal of corepressor complexes required for gene activation. As a result, NCoR complexes are not cleared from the promoter and target genes are maintained in a repressed state. This mechanism provides an explanation for how an agonist-bound nuclear receptor can be converted from an activator of transcription to a promoter-specific repressor of NF-kappaB target genes that regulate immunity and homeostasis.
- Published
- 2005
- Full Text
- View/download PDF
34. Controlling nuclear receptors: the circular logic of cofactor cycles.
- Author
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Perissi V and Rosenfeld MG
- Subjects
- Animals, Gene Expression Regulation, Homeostasis, Humans, Mammals, Models, Molecular, Promoter Regions, Genetic, Protein Processing, Post-Translational, Receptors, Cytoplasmic and Nuclear genetics, Receptors, Cytoplasmic and Nuclear metabolism, Transcription, Genetic, Receptors, Cytoplasmic and Nuclear physiology
- Abstract
Nuclear receptors regulate many biologically important processes in development and homeostasis by their bimodal function as repressors and activators of gene transcription. A finely tuned modulation of the transcriptional activities of nuclear receptors is crucial for determining highly specific and diversified programmes of gene expression. Recent studies have provided insights into the molecular mechanisms that are required to switch between repression and activation functions, the combinatorial roles of the multiple cofactor complexes that are required for mediating transcriptional regulation, and the central question of how several different signalling pathways can be integrated at the nuclear level to achieve specific profiles of gene expression.
- Published
- 2005
- Full Text
- View/download PDF
35. A nuclear receptor corepressor transcriptional checkpoint controlling activator protein 1-dependent gene networks required for macrophage activation.
- Author
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Ogawa S, Lozach J, Jepsen K, Sawka-Verhelle D, Perissi V, Sasik R, Rose DW, Johnson RS, Rosenfeld MG, and Glass CK
- Subjects
- Animals, Cells, Cultured, Genes, jun, Macrophage Activation genetics, Mice, Mice, Knockout, Models, Biological, Nuclear Proteins deficiency, Nuclear Proteins genetics, Nuclear Receptor Co-Repressor 1, Recombinant Proteins genetics, Recombinant Proteins metabolism, Repressor Proteins genetics, Signal Transduction, Transcription Factor AP-1 genetics, Transcription, Genetic, Transfection, Macrophage Activation physiology, Nuclear Proteins metabolism, Repressor Proteins metabolism, Transcription Factor AP-1 metabolism
- Abstract
The nuclear receptor corepressor (NCoR) and the related factor known as silencing mediator of retinoic acid and thyroid hormone receptor (SMRT) are essential components of multiprotein complexes that mediate active repression by unliganded nuclear receptors. Recent studies suggest that NCoR and SMRT can interact with and exert repressive effects on several other classes of DNA-binding transcription factors, but the physiological importance of these interactions has not been established. Here, investigation of endogenous transcriptional programs regulated by NCoR in macrophages reveals that NCoR acts as a transcriptional checkpoint for activator protein (AP)-1-dependent gene networks that regulate diverse biological processes including inflammation, cell migration, and collagen catabolism, with loss of NCoR, resulting in derepression of AP-1 target genes. The NCoR corepressor complex imposes an active block of exchange of c-Jun for c-Jun/c-Fos heterodimers, with targeted deletion of the c-Jun locus, resulting in loss of NCoR complexes from AP-1 target genes under basal conditions. The checkpoint function of NCoR is relieved by signal-dependent phosphorylation of c-Jun, which directs removal of NCoR/HDAC3/TBL1/TBLR1 complexes through recruitment of a specific ubiquitylation complex, as a prerequisite to the default binding of c-Jun/c-Fos heterodimers and transcriptional activation. The requirement for a checkpoint function to achieve the appropriate dynamic range of transcriptional responses to inflammatory signals is likely to be used by other signal-dependent transcription factors that regulate diverse homeostatic and developmental processes.
- Published
- 2004
- Full Text
- View/download PDF
36. A corepressor/coactivator exchange complex required for transcriptional activation by nuclear receptors and other regulated transcription factors.
- Author
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Perissi V, Aggarwal A, Glass CK, Rose DW, and Rosenfeld MG
- Subjects
- Adipocytes cytology, Animals, Cell Differentiation, Cells, Cultured, Cysteine Endopeptidases metabolism, Embryo, Mammalian cytology, Endothelium, Vascular cytology, Gene Deletion, Genetic Vectors, Ligands, Mice, Microscopy, Fluorescence, Models, Biological, Models, Genetic, Multienzyme Complexes metabolism, NF-kappa B metabolism, Neurons cytology, Nuclear Proteins metabolism, Precipitin Tests, Proteasome Endopeptidase Complex, Protein Binding, Proto-Oncogene Proteins c-jun metabolism, RNA chemistry, Receptors, Cytoplasmic and Nuclear metabolism, Repressor Proteins metabolism, Reverse Transcriptase Polymerase Chain Reaction, Signal Transduction, Stem Cells metabolism, Transcription Factors metabolism, Transducin metabolism, Ubiquitin metabolism, Cell Nucleus metabolism, Transcriptional Activation
- Abstract
The mechanisms that control the precisely regulated switch from gene repression to gene activation represent a central question in mammalian development. Here, we report that transcriptional activation mediated by liganded nuclear receptors unexpectedly requires the actions of two highly related F box/WD-40-containing factors, TBL1 and TBLR1, initially identified as components of an N-CoR corepressor complex. TBL1/TBLR1 serve as specific adaptors for the recruitment of the ubiquitin conjugating/19S proteasome complex, with TBLR1 selectively serving to mediate a required exchange of the nuclear receptor corepressors, N-CoR and SMRT, for coactivators upon ligand binding. Tbl1 gene deletion in embryonic stem cells severely impairs PPARgamma-induced adipogenic differentiation, indicating that TBL1 function is also biologically indispensable for specific nuclear receptor-mediated gene activation events. The role of TBLR1 and TBL1 in cofactor exchange appears to also operate for c-Jun and NFkappaB and is therefore likely to be prototypic of similar mechanisms for other signal-dependent transcription factors.
- Published
- 2004
- Full Text
- View/download PDF
37. Tissue-specific regulation of retinal and pituitary precursor cell proliferation.
- Author
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Li X, Perissi V, Liu F, Rose DW, and Rosenfeld MG
- Subjects
- Animals, Apoptosis, Cell Cycle, Cell Cycle Proteins genetics, Cell Cycle Proteins metabolism, Cell Line, Cyclin-Dependent Kinase Inhibitor p27, Cyclin-Dependent Kinases antagonists & inhibitors, DNA-Binding Proteins, Embryo, Mammalian cytology, Eye Proteins metabolism, Mice, Nuclear Proteins metabolism, Organ Specificity, Pituitary Gland embryology, Promoter Regions, Genetic, Proto-Oncogene Proteins genetics, Proto-Oncogene Proteins metabolism, Recombinant Fusion Proteins metabolism, Repressor Proteins metabolism, Retina embryology, Retinal Ganglion Cells cytology, Retinal Ganglion Cells physiology, Transcription Factors, Transcription, Genetic, Transfection, Tumor Suppressor Proteins genetics, Tumor Suppressor Proteins metabolism, Up-Regulation, Cell Division, Homeodomain Proteins genetics, Homeodomain Proteins metabolism, Pituitary Gland cytology, Retina cytology, Stem Cells physiology, Trans-Activators genetics, Trans-Activators metabolism
- Abstract
Mammalian organogenesis requires the expansion of pluripotent precursor cells before the subsequent determination of specific cell types, but the tissue-specific molecular mechanisms that regulate the initial expansion of primordial cells remain poorly defined. We have genetically established that Six6 homeodomain factor, acting as a strong tissue-specific repressor, regulates early progenitor cell proliferation during mammalian retinogenesis and pituitary development. Six6, in association with Dach corepressors, regulates proliferation by directly repressing cyclin-dependent kinase inhibitors, including the p27Kip1 promoter. These data reveal a molecular mechanism by which a tissue-specific transcriptional repressor-corepressor complex can provide an organ-specific strategy for physiological expansion of precursor populations.
- Published
- 2002
- Full Text
- View/download PDF
38. The histone deacetylase-3 complex contains nuclear receptor corepressors.
- Author
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Wen YD, Perissi V, Staszewski LM, Yang WM, Krones A, Glass CK, Rosenfeld MG, and Seto E
- Subjects
- Acetylation, Amino Acid Sequence, HeLa Cells, Histone Deacetylases genetics, Humans, Molecular Sequence Data, Receptors, Cytoplasmic and Nuclear genetics, Signal Transduction, Histone Deacetylases metabolism, Receptors, Cytoplasmic and Nuclear metabolism
- Abstract
Acetylation and deacetylation of nucleosomal histones have profound effects on gene transcription in all eukaryotes. In humans, three highly homologous class I and four class II histone deacetylase (HDAC) enzymes have been identified to date. The class I deacetylases HDAC1 and HDAC2 are components of multisubunit complexes, one of which could associate with the nuclear hormone receptor corepressor, N-CoR. N-CoR also interacts with class II deacetylases HDAC4, HDAC5, and HDAC7. In comparison with HDAC1 and HDAC2, HDAC3 remains relatively uncharacterized, and very few proteins have been shown to interact with HDAC3. Using an affinity purification approach, we isolated an enzymatically active HDAC3 complex that contained members of the nuclear receptor corepressor family. Deletion analysis of N-CoR revealed that HDAC3 binds multiple N-CoR regions in vitro and that all of these regions are required for maximal binding in vivo. The N-CoR domains that interact with HDAC3 are distinct from those that bind other HDACs. Transient overexpression of HDAC3 and microinjection of Abs against HDAC3 showed that a component of transcriptional repression mediated by N-CoR depends on HDAC3. Interestingly, data suggest that interaction with a region of N-CoR augments the deacetylase activity of HDAC3. These results provide a possible molecular mechanism for HDAC3 regulation and argue that N-CoR is a platform in which distinct domains can interact with most of the known HDACs.
- Published
- 2000
- Full Text
- View/download PDF
39. AP-2 transcription factors in the regulation of ERBB2 gene transcription by oestrogen.
- Author
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Perissi V, Menini N, Cottone E, Capello D, Sacco M, Montaldo F, and De Bortoli M
- Subjects
- Breast Neoplasms, DNA Footprinting, DNA-Binding Proteins antagonists & inhibitors, DNA-Binding Proteins biosynthesis, DNA-Binding Proteins metabolism, Down-Regulation genetics, Estrogens metabolism, Humans, Promoter Regions, Genetic genetics, Promoter Regions, Genetic physiology, Receptors, Estrogen genetics, Transcription Factor AP-2, Transcription Factors biosynthesis, Transcription Factors metabolism, Tumor Cells, Cultured, DNA-Binding Proteins physiology, Estrogens physiology, Genes, erbB-2 physiology, Transcription Factors physiology, Transcription, Genetic genetics
- Abstract
Transcription of the ERBB2 oncogene is repressed by oestrogen in human breast cancer cells. We show that a 218 bp fragment of the human ERBB2 gene promoter is responsive to oestrogen in transient transfection in ZR75.1 and SKBR.3 cells when the oestrogen receptor is expressed. Deletion analysis of this fragment shows that a sequence located at the 5' end, which is known to mediate ERBB2 overexpression in breast cancer, is also responsible for the oestrogen response. This sequence binds AP-2 transcription factors and appears functionally identical to an element of the oestrogen-dependent enhancer described in the first intron of human ERBB2. We observed that oestrogen treatment down-regulates expression of AP-2 proteins but does not affect the DNA binding activity of AP-2. Constitutive expression of AP-2beta or AP-2gamma, but not AP-2alpha, abrogates the estrogenic repression. Our results demonstrate that AP-2 transcription factors are implicated in the oestrogenic regulation of ERBB2 gene expression and suggest a complex interplay involving the different AP-2 isoforms and other unidentified factors.
- Published
- 2000
- Full Text
- View/download PDF
40. Factor-specific modulation of CREB-binding protein acetyltransferase activity.
- Author
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Perissi V, Dasen JS, Kurokawa R, Wang Z, Korzus E, Rose DW, Glass CK, and Rosenfeld MG
- Subjects
- Acetylation, Animals, Binding Sites, CREB-Binding Protein, Cell Line, Cyclic AMP Response Element-Binding Protein metabolism, Fibroblasts, Histones metabolism, Kinetics, Rats, Recombinant Fusion Proteins biosynthesis, Recombinant Proteins metabolism, Substrate Specificity, Transfection, Acetyltransferases metabolism, Adenovirus E1A Proteins metabolism, Nuclear Proteins metabolism, Trans-Activators metabolism
- Abstract
CREB-binding proteins (CBP) and p300 are essential transcriptional coactivators for a large number of regulated DNA-binding transcription factors, including CREB, nuclear receptors, and STATs. CBP and p300 function in part by mediating the assembly of multiprotein complexes that contain additional cofactors such as p300/CBP interacting protein (p/CIP), a member of the p160/SRC family of coactivators, and the p300/CBP associated factor p/CAF. In addition to serving as molecular scaffolds, CBP and p300 each possess intrinsic acetyltransferase activities that are required for their function as coactivators. Here we report that the adenovirus E1A protein inhibits the acetyltransferase activity of CBP on binding to the C/H3 domain, whereas binding of CREB, or a CREB/E1A fusion protein to the KIX domain, fails to inhibit CBP acetyltransferase activity. Surprisingly, p/CIP can either inhibit or stimulate CBP acetyltransferase activity depending on the specific substrate evaluated and the functional domains present in the p/CIP protein. While the CBP interaction domain of p/CIP inhibits acetylation of histones H3, H4, or high mobility group by CBP, it enhances acetylation of other substrates, such as Pit-1. These observations suggest that the acetyltransferase activities of CBP/p300 and p/CAF can be differentially modulated by factors binding to distinct regions of CBP/p300. Because these interactions are likely to result in differential effects on the coactivator functions of CBP/p300 for different classes of transcription factors, regulation of CBP/p300 acetyltransferase activity may represent a mechanism for integration of diverse signaling pathways.
- Published
- 1999
- Full Text
- View/download PDF
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