9 results on '"Zheng, Gen"'
Search Results
2. Flow-dependent epigenetic regulation of IGFBP5 expression by H3K27me3 contributes to endothelial anti-inflammatory effects
- Author
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Shuya Zhang, Suowen Xu, Shuyi Si, Marina Koroleva, Jaroslav Pelisek, Zheng Gen Jin, Yanni Xu, Meimei Yin, Peng Liu, and Peter J. Little
- Subjects
0301 basic medicine ,Endothelium ,H3K27me3 ,Anti-Inflammatory Agents ,Medicine (miscellaneous) ,macromolecular substances ,Methylation ,Epigenesis, Genetic ,Histones ,Mice ,03 medical and health sciences ,Downregulation and upregulation ,Gene expression ,Human Umbilical Vein Endothelial Cells ,medicine ,Animals ,Humans ,Enhancer of Zeste Homolog 2 Protein ,EZH2 ,Epigenetics ,RNA, Small Interfering ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,biology ,Polycomb Repressive Complex 2 ,Genetic Therapy ,Atherosclerosis ,endothelial cells ,3. Good health ,Chromatin ,Cell biology ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,Histone ,medicine.anatomical_structure ,biology.protein ,IGFBP5 ,Insulin-Like Growth Factor Binding Protein 5 ,PRC2 ,epigenetic ,Research Paper - Abstract
Rationale: Atherosclerosis is a chronic inflammatory and epigenetic disease that is influenced by different patterns of blood flow. However, the epigenetic mechanism whereby atheroprotective flow controls endothelial gene programming remains elusive. Here, we investigated the possibility that flow alters endothelial gene expression through epigenetic mechanisms. Methods: En face staining and western blot were used to detect protein expression. Real-time PCR was used to determine relative gene expression. RNA-sequencing of human umbilical vein endothelial cells treated with siRNA of enhancer of zeste homolog 2 (EZH2) or laminar flow was used for transcriptional profiling. Results: We found that trimethylation of histone 3 lysine 27 (H3K27me3), a repressive epigenetic mark that orchestrates gene repression, was reduced in laminar flow areas of mouse aorta and flow-treated human endothelial cells. The decrease of H3K27me3 paralleled a reduction in the epigenetic “writer”-EZH2, the catalytic subunit of the polycomb repressive complex 2 (PRC2). Moreover, laminar flow decreased expression of EZH2 via mechanosensitive miR101. Genome-wide transcriptome profiling studies in endothelial cells treated with EZH2 siRNA and flow revealed the upregulation of novel mechanosensitive gene IGFBP5 (insulin-like growth factor-binding protein 5), which is epigenetically silenced by H3K27me3. Functionally, inhibition of H3K27me3 by EZH2 siRNA or GSK126 (a specific EZH2 inhibitor) reduced H3K27me3 levels and monocyte adhesion to endothelial cells. Adenoviral overexpression of IGFBP5 also recapitulated the anti-inflammatory effects of H3K27me3 inhibition. More importantly, we observed EZH2 upregulation, and IGFBP5 downregulation, in advanced atherosclerotic plaques from human patients. Conclusion: Taken together, our findings reveal that atheroprotective flow reduces H3K27me3 as a chromatin-based mechanism to augment the expression of genes that confer an anti-inflammatory response in the endothelium. Our study exemplifies flow-dependent epigenetic regulation of endothelial gene expression, and also suggests that targeting the EZH2/H3K27me3/IGFBP5 pathway may offer novel therapeutics for inflammatory disorders such as atherosclerosis.
- Published
- 2018
3. SENCR stabilizes vascular endothelial cell adherens junctions through interaction with CKAP4.
- Author
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Qing Lyu, Suowen Xu, Yuyan Lyu, Mihyun Choi, Christie, Christine K., Slivano, Orazio J., Rahman, Arshad, Zheng-Gen Jin, Xiaochun Long, Yawei Xu, and Miano, Joseph M.
- Subjects
LINCRNA ,VASCULAR smooth muscle ,ENDOTHELIAL cells ,GENE expression ,CARRIER proteins ,SHEAR strength ,IMMUNOPRECIPITATION - Abstract
SENCR is a human-specific, vascular cell-enriched long-noncoding RNA (lncRNA) that regulates vascular smooth muscle cell and endothelial cell (EC) phenotypes. The underlying mechanisms of action of SENCR in these and other cell types is unknown. Here, levels of SENCR RNA are shown to be elevated in several differentiated human EC lineages subjected to laminar shear stress. Increases in SENCR RNA are also observed in the laminar shear stress region of the adult aorta of humanized SENCR-expressing mice, but not in disturbed shear stress regions. SENCR loss-of-function studies disclose perturbations in EC membrane integrity resulting in increased EC permeability. Biotinylated RNA pull-down and mass spectrometry establish an abundant SENCR-binding protein, cytoskeletal-associated protein 4 (CKAP4); this ribonucleoprotein complex was further confirmed in an RNA immunoprecipitation experiment using an antibody to CKAP4. Structure-function studies demonstrate a noncanonical RNA-binding domain in CKAP4 that binds SENCR. Upon SENCR knockdown, increasing levels of CKAP4 protein are detected in the EC surface fraction. Furthermore, an interaction between CKAP4 and CDH5 is enhanced in SENCR-depleted EC. This heightened association appears to destabilize the CDH5/CTNND1 complex and augment CDH5 internalization, resulting in impaired adherens junctions. These findings support SENCR as a flow-responsive lncRNA that promotes EC adherens junction integrity through physical association with CKAP4, thereby stabilizing cell membrane-bound CDH5. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
4. PKA phosphorylates histone deacetylase 5 and prevents its nuclear export, leading to the inhibition of gene transcription and cardiomyocyte hypertrophy
- Author
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Chelsea Wong, Jinjing Zhao, Ji Young Kim, Chang Hoon Ha, Bong Sook Jhun, Weiye Wang, and Zheng Gen Jin
- Subjects
Transcription, Genetic ,Green Fluorescent Proteins ,Immunoblotting ,Molecular Sequence Data ,Active Transport, Cell Nucleus ,Histone Deacetylases ,Substrate Specificity ,Gene expression ,Chlorocebus aethiops ,medicine ,Cyclic AMP ,Animals ,Humans ,Myocytes, Cardiac ,Amino Acid Sequence ,Phosphorylation ,Protein kinase A ,Nuclear export signal ,Cell Shape ,Cells, Cultured ,Cell Nucleus ,Histone deacetylase 5 ,Cyclic AMP-Dependent Protein Kinase Catalytic Subunits ,Multidisciplinary ,biology ,Sequence Homology, Amino Acid ,Kinase ,Colforsin ,Biological Sciences ,Rats ,Cell nucleus ,Histone ,medicine.anatomical_structure ,Biochemistry ,Animals, Newborn ,Microscopy, Fluorescence ,COS Cells ,biology.protein - Abstract
Dynamic nucleocytoplasmic shuttling of class IIa histone deacetylases (HDACs) is a fundamental mechanism regulating gene transcription. Recent studies have identified several protein kinases that phosphorylate HDAC5, leading to its exportation from the nucleus. However, the negative regulatory mechanisms for HDAC5 nuclear exclusion remain largely unknown. Here we show that cAMP-activated protein kinase A (PKA) specifically phosphorylates HDAC5 and prevents its export from the nucleus, leading to suppression of gene transcription. PKA interacts directly with HDAC5 and phosphorylates HDAC5 at serine 280, an evolutionarily conserved site. Phosphorylation of HDAC5 by PKA interrupts the association of HDAC5 with protein chaperone 14-3-3 and hence inhibits stress signal-induced nuclear export of HDAC5. An HDAC5 mutant that mimics PKA-dependent phosphorylation localizes in the nucleus and acts as a dominant inhibitor for myocyte enhancer factor 2 transcriptional activity. Molecular manipulations of HDAC5 show that PKA-phosphorylated HDAC5 inhibits cardiac fetal gene expression and cardiomyocyte hypertrophy. Our findings identify HDAC5 as a substrate of PKA and reveal a cAMP/PKA-dependent pathway that controls HDAC5 nucleocytoplasmic shuttling and represses gene transcription. This pathway may represent a mechanism by which cAMP/PKA signaling modulates a wide range of biological functions and human diseases such as cardiomyopathy.
- Published
- 2010
5. Improvement of Transmembrane Transport Mechanism Study of Imperatorin on P-Glycoprotein-Mediated Drug Transport.
- Author
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Zheng-Gen Liao, Tao Tang, Xue-Jing Guan, Wei Dong, Jing Zhang, Guo-Wei Zhao, Ming Yang, and Xin-Li Liang
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MEMBRANE proteins , *P-glycoprotein , *ADENOSINE triphosphatase , *WESTERN immunoblotting , *REVERSE transcriptase polymerase chain reaction , *VERAPAMIL , *GENE expression , *MULTIDRUG resistance - Abstract
P-glycoprotein (P-gp) affects the transport of many drugs; including puerarin and vincristine. Our previous study demonstrated that imperatorin increased the intestinal absorption of puerarin and vincristine by inhibiting P-gp-mediated drug efflux. However; the underlying mechanism was not known. The present study investigated the mechanism by which imperatorin promotes P-gp-mediated drug transport. We used molecular docking to predict the binding force between imperatorin and P-gp and the effect of imperatorin on P-gp activity. P-gp efflux activity and P-gp ATPase activity were measured using a rhodamine 123 (Rh-123) accumulation assay and a Pgp-Glo™ assay; respectively. The fluorescent probe 1,6-diphenyl-1,3,5-hexatriene (DPH) was used to assess cellular membrane fluidity in MDCK-MDR1 cells. Western blotting was used to analyze the effect of imperatorin on P-gp expression; and P-gp mRNA levels were assessed by qRT-PCR. Molecular docking results demonstrated that the binding force between imperatorin and P-gp was much weaker than the force between P-gp and verapamil (a P-gp substrate). Imperatorin activated P-gp ATPase activity; which had a role in the inhibition of P-gp activity. Imperatorin promoted Rh-123 accumulation in MDCK-MDR1 cells and decreased cellular membrane fluidity. Western blotting demonstrated that imperatorin inhibited P-gp expression; and qRT-PCR revealed that imperatorin down-regulated P-gp (MDR1) gene expression. Imperatorin decreased P-gp-mediated drug efflux by inhibiting P-gp activity and the expression of P-gp mRNA and protein. Our results suggest that imperatorin could down-regulate P-gp expression to overcome multidrug resistance in tumors. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
6. PKA phosphorylates histone deacetylase 5 and prevents its nuclear export, leading to the inhibition of gene transcription and cardiomyocyte hypertrophy.
- Author
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Chang Hoon Ha, Ji Young Kim, Jinjing Zhao, Weiye Wang, Bong Sook Jhun, Chelsea Wong, and Zheng Gen Jin
- Subjects
GENE expression ,PROTEIN kinases ,PHOSPHORYLATION ,HYPERTROPHY ,CARDIOMYOPATHIES ,GENETIC transcription ,HEART cells - Abstract
Dynamic nucleocytoplasmic shuttling of class IIa histone deacetylases (HDACs) is a fundamental mechanism regulating gene transcription. Recent studies have identified several protein kinases that phosphorylate HDAC5, leading to its exportation from the nucleus. However, the negative regulatory mechanisms for HDAC5 nuclear exclusion remain largely unknown. Here we show that cAMP-activated protein kinase A (PKA) specifically phosphorylates HDAC5 and prevents its export from the nucleus, leading to suppression of gene transcription. PKA interacts directly with HDAC5 and phosphorylates HDAC5 at serine 280, an evolutionarily conserved site. Phosphorylation of HDAC5 by PKA interrupts the association of HDAC5 with protein chaperone 14-3-3 and hence inhibits stress signal-induced nuclear export of HDAC5. An HDAC5 mutant that mimics PKA-dependent phosphorylation localizes in the nucleus and acts as a dominant inhibitor for myocyte enhancer factor 2 transcriptional activity. Molecular manipulations of HDAC5 show that PKA-phosphorylated HDAC5 inhibits cardiac fetal gene expression and cardiomyocyte hypertrophy. Our findings identify HDAC5 as a substrate of PKA and reveal a cAMP/PKA-dependent pathway that controls HDAC5 nucleocytoplasmic shuttling and represses gene transcription. This pathway may represent a mechanism by which cAMP/PKA signaling modulates a wide range of biological functions and human diseases such as cardiomyopathy. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
7. Protein kinase D controls voluntary-running-induced skeletal muscle remodelling.
- Author
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Kornelia Ellwanger, Christine Kienzle, Sylke Lutz, Zheng‑Gen Jin, Maria T. Wiekowski, Klaus Pfizenmaier, and Angelika Hausser
- Subjects
PROTEIN kinases ,SKELETAL muscle ,HISTONE deacetylase ,GENE expression ,TRANSCRIPTION factors ,LABORATORY mice ,CELLULAR signal transduction ,PROMOTERS (Genetics) - Abstract
Skeletal muscle responds to exercise by activation of signalling pathways that co-ordinate gene expression to sustain muscle performance. MEF2 (myocyte enhancer factor 2)-dependent transcriptional activation of MHC (myosin heavy chain) genes promotes the transformation from fast-twitch into slow-twitch fibres, with MEF2 activity being tightly regulated by interaction with class IIa HDACs (histone deacetylases). PKD (protein kinase D) is known to directly phosphorylate skeletal muscle class IIa HDACs, mediating their nuclear export and thus derepression of MEF2. In the present study, we report the generation of transgenic mice with inducible conditional expression of a dominant-negative PKD1kd (kinase-dead PKD1) protein in skeletal muscle to assess the role of PKD in muscle function. In control mice, long-term voluntary running experiments resulted in a switch from type IIb+IId/x to type IIa plantaris muscle fibres as measured by indirect immunofluorescence of MHCs isoforms. In mice expressing PKD1kd, this fibre type switch was significantly impaired. These mice exhibited altered muscle fibre composition and decreased running performance compared with control mice. Our findings thus indicate that PKD activity is essential for exercise-induced MEF2-dependent skeletal muscle remodelling in vivo. [ABSTRACT FROM AUTHOR]
- Published
- 2011
- Full Text
- View/download PDF
8. Expression, regulation and function of miR-126 in the mouse choroid vasculature.
- Author
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Zhao, Fangkun, Anderson, Chastain, Karnes, Sara, Zhou, Qinbo, Ma, Jing, Jin, Zheng-Gen, Bhattacharjee, Partha S., and Wang, Shusheng
- Subjects
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CHOROID , *BLOOD vessels , *MICRORNA , *GENE expression , *GENETIC regulation , *NEOVASCULARIZATION - Abstract
MicroRNA miR-126 has been shown to be required for proper angiogenesis in several models. However, its expression, regulation and function in the mouse choroid remain unclear. Our previous data has shown that miR-126 expression is enriched in the endothelial cells (ECs) of the mouse choroid. Here we report that a 5.5 kb Egfl7/miR-126 promoter drives the expression of miR-126 in the choroid ECs during choroidal vascular development. The expression of miR-126 in the ECs is regulated by flow stress likely through Krüppel-like transcriptional factors. miR-126 −/− mice show mildly delayed choroidal vascular development, but adult knockout mice develop periphery choroidal vascular lesions. This study suggests that miR-126 is largely dispensable for mouse choroidal development but required for maintaining choroidal vasculature integrity. [ABSTRACT FROM AUTHOR]
- Published
- 2018
- Full Text
- View/download PDF
9. Protein Kinase D-dependent Phosphorylation and Nuclear Export of Histone Deacetylase 5 Mediates Vascular Endothelial Growth Factor-induced Gene Expression and Angiogenesis.
- Author
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Chang Hoon Ha, Weiye Wang, Bong Sook Jhun, Chelsea Wong, Hausser, Angelika, Pfizenmaier, Klaus, McKinsey, Timothy A., Olson, Eric N., and Zheng-Gen Jin
- Subjects
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PROTEIN kinases , *HISTONE deacetylase , *PHOSPHORYLATION , *VASCULAR endothelial growth factors , *GENE expression , *NEOVASCULARIZATION - Abstract
Vascular endothelial growth factor (VEGF) is essential for normal and pathological angiogenesis. However, the signaling pathways linked to gene regulation in VEG F-induced angiogenesis are not fully understood. Here we demonstrate a critical role of protein kinase D (PKD) and histone deacetylase 5 (HDAC5) in VEGF-induced gene expression and angiogenesis. We found that VEGF stimulated HDAC5 phosphorylation and nuclear export in endothelial cells through a VEGF receptor 2-phospholipase Cγ-protein kinase C-PKD-dependent pathway. We further showed that the PKD-HDAC5 pathway mediated myocyte enhancer factor-2 transcriptional activation and a specific subset of gene expression in response to VEGF, including NR4A1, an orphan nuclear receptor involved in angiogenesis. Specifically, inhibition of PKD by overexpression of the PKD kinase-negative mutant prevents VEGF-induced HDAC5 phosphorylation and nuclear export as well as NR4A1 induction. Moreover, a mutant of HDAC5 specifically deficient in PKD-dependent phosphorylation inhibited VEGF-mediated NR4A1 expression, endothelial cell migration, and in vitro angiogenesis. These findings suggest that the PKD-HDAC5 pathway plays an important role in VEGF regulation of gene transcription and angiogenesis. [ABSTRACT FROM AUTHOR]
- Published
- 2008
- Full Text
- View/download PDF
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