王文恭
个性化签名
- 姓名:王文恭
- 目前身份:
- 担任导师情况:
- 学位:
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学术头衔:
博士生导师
- 职称:-
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学科领域:
生物化学
- 研究兴趣:
王文恭,男,1963年3月出生。教授,博士生导师。
学习与工作经历:
1979-1984山西医科大学医学系,学士学位
1984-1988山西长治医学院,助教
1988-1991中山医科大学,硕士学位
1991-1993山西长治医学院,讲师
1993-1996北京医科大学,博士学位
1997-2004National Institute on Aging, NIH, Maryland
2004-2005Assistant Professor, Center for Vascular Biology, University of Connecticut Health Center
2005-北京大学医学部生物化学与分子生物学系,教授
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1260
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关注数
0
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成果阅读
377
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成果数
8
【期刊论文】HuR Regulates p21 mRNA Stabilization by UV Light
王文恭, WENGONG WANG, HENRY FURNEAUX, HUIMING CHENG, M. CRAIG CALDWELL, DOROTHY HUTTER, YUSEN LIU, NIKKI HOLBROOK, AND MYRIAM GOROSPE *
MOLECULAR AND CELLULAR BIOLOGY, Feb. 2000, p. 760-769,-0001,():
-1年11月30日
Expression of the cyclin-dependent kinase inhibitor p21 is highly induced by many stresses, including exposure to short-wavelength UV light (UVC), which increases p21 mRNA stability. Investigation into the mechanisms underlying this stabilization process revealed that proteins present in cytoplasmic lysates of human RKO colorectal carcinoma cells formed complexes with p21 mRNA that were inducible by treatment with UVC and other stress agents. The ubiquitous Elav-type RNA-binding protein HuR was identified within the p21 mRNA-protein complexes, as antibodies recognizing HuR supershifted these complexes and revealed HuR-immunoreactive proteins complexing with p21 mRNA on Western blots. Lowering of endogenous HuR levels through expression of antisense HuR decreased p21 RNA-protein complexes, greatly reduced the UVC inducibility and half-life of p21 mRNA, and prevented UVC-mediated induction of luciferase activity in p21 3* untranslated region-containing reporter constructs. Our findings indicate that HuR plays a major role in regulating stress-induced p21 expression by enhancing p21 mRNA stability and that these effects are coupled to HuR’s elevated presence in the cytoplasm.
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【期刊论文】HuR regulates cyclin A and cyclin B1 mRNA stability during cell proliferation
王文恭, Wengong Wang, M.Craig Caldwell, Shankung Lin, Henry Furneaux and Myriam Gorospe
The EMBO Journal Vol. 19 No.10 pp. 2340-2350, 2000,-0001,():
-1年11月30日
Colorectal carcinoma RKO cells expressing reduced levels of the RNA-binding protein HuR (ASHuR) displayed markedly reduced growth. In synchronous RKO populations, HuR was almost exclusively nuclear during early G1, increasing in the cytoplasm during late G1, S and G2. The expression and half-life of mRNAs encoding cyclins A and BI similarly increased during S and G2, then declined, indicating that mRNA stabilization contributed to their cell cycle-regulated expression. In gel-shift assays using radlolabeled cyclin RNA transcripts and RKO protein extracts, only those transcripts corresponding to the 3'-untranslated regions of cyclins A and BI formed RNA-protein complexes in a cell cycle-dependent fashion. HuR directly bound mRNAs encoding cyclins A and BI, as anti-HuR antibodies supershifted such RNA-protein complexes. Importantly, the expression and half-life of mRNAs encoding cyclins A and BI were reduced in ASHuR RKO cells. Our results indicate that HuR may play a critical role in cell proliferation, at least in part by mediating cell cycledependent stabilization of mRNAs encoding cyclins A and B1.
cyclin A, cyclin B1, HaR, proliferation, mRNA stability
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王文恭, SHANKUNG LIN, WENGONG WANG, GERALD M. WILSON, XIAOLING YANG, GARY BREWER, NIKKI J. HOLBROOK, AND MYRIAM GOROSPE *
MOLECULAR AND CELLULAR BIOLOGY, Nov. 2000, p. 7903-7913,-0001,():
-1年11月30日
Prostaglandin A2 (PGA2), an experimental chemotherapeutic agent, causes growth arrest associated with decreased cyclin D1 expression in several cancer cell lines. Here, using human non-small-cell lung carcinoma H1299 cells, we investigated the mechanisms whereby PGA2 down-regulates cyclin D1 expression. Transcription rates of the cyclin D1 gene, studied using a cyclin D1 promoter-luciferase construct and nuclear run-on assays, were not affected by PGA2 treatment. Instead, the cyclin D1 mRNA was rendered unstable after exposure to PGA2. Since the stability of labile mRNA is modulated through binding of proteins to specific mRNA sequences, we sought to identify protein(s) recognizing the cyclin D1 mRNA. In electrophoretic mobility-shift assays using radiolabeled RNA probes derived from different regions of cyclin D1 mRNA, we observed that (i) lysates prepared from PGA2-treated cells exhibited enhanced protein-cyclin D1 RNA complex formation; (ii) the kinetics of complex formation correlated closely with that of cyclin D1 mRNA loss; and (iii) binding occurred within a 390-base cyclin D1 3* untranslated region (UTR) (K12). This binding activity could be cross-linked, revealing proteins ranging from 30 to 47 kDa. The RNA-binding protein AUF1, previously associated with the degradation of target mRNAs, bound cyclin D1 mRNA, because anti-AUF1 antibodies were capable of supershifting or immunoprecipitating cyclin D1 mRNA-protein complexes. Finally, insertion of K12 in the 3*UTR of reporter genes markedly reduced the expression and half-life of the resulting chimeric mRNAs in transfected, PGA2-treated cells. Our data demonstrate that PGA2 down-regulates cyclin D1 expression by decreasing cyclin D1 mRNA stability and implicates a 390-base element in the 3*UTR in this regulation.
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王文恭, WENGONG WANG, XIAOLING YANG, VINCENT J. CRISTOFALO, NIKKI J. HOLBROOK, AND MYRIAM GOROSPE *
MOLECULAR AND CELLULAR BIOLOGY, Sept. 2001, p. 5889-5898,-0001,():
-1年11月30日
Cellular aging is accompanied by alterations in gene expression patterns. Here, using two models of replicative senescence, we describe the influence of the RNA-binding protein HuR in regulating the expression of several genes whose expression decreases during senescence. We demonstrate that HuR levels, HuR binding to target mRNAs encoding proliferative genes, and the half-lives of such mRNAs are lower in senescent cells. Importantly, overexpression of HuR in senescent cells restored a "younger" phenotype, while a reduction in HuR expression accentuated the senescent phenotype. Our studies highlight a critical role for HuR during the process of replicative senescence.
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【期刊论文】AMP-Activated Kinase Regulates Cytoplasmic HuR
王文恭, Wengong Wang, Jinshui Fan, Xiaoling Yang, Stefanie Fürer-Galban, Isabel Lopez de Silanes, Cayetano von Kobbe, Jia Guo, Steve N. Georas, Fabienne Foufelle, D. Grahame Hardie, David Carling, and Myriam Gorospe *
MOLECULAR AND CELLULAR BIOLOGY, May 2002, p. 3425-3436,-0001,():
-1年11月30日
While transport of RNA-binding protein HuR from nucleus to cytoplasm is emerging as a key regulatory step for HuR function, the mechanisms underlying this process remain poorly understood. Here, we report that the AMP-activated kinase (AMPK), an enzyme involved in responding to metabolic stresses, potently regulates the levels of cytoplasmic HuR. Inhibition of AMPK, accomplished either through cell treatment or by adenovirus infection to express dominant-negative AMPK, was found to increase the level of HuR in the cytoplasm and to enhance the binding of HuR to p21, cyclin B1, and cyclin A mRNA transcripts and elevate their expression and half-lives. Conversely, AMPK activation, achieved by means including infection to express constitutively active AMPK, resulted in reduced cytoplasmic HuR; decreased levels and half-lives of mRNAs encoding p21, cyclin A, and cyclin B1; and diminished HuR association with the corresponding transcripts. We therefore propose a novel function for AMPK as a regulator of cytoplasmic HuR levels, which in turn influences the mRNAstabilizing function of HuR and the expression of HuR target transcripts.
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【期刊论文】Global analysis of stress-regulated mRNA turnover by using cDNA arrays
王文恭, Jinshui Fan*, Xiaoling Yang*, Wengong Wang*, William H. Wood 〣†, Kevin G. Becker†, and Myriam Gorospe*‡
PNASㄧAugust 6, 2002ㄧvol. 99ㄧno.16ㄧ10611-10616,-0001,():
-1年11月30日
cDNA array technology has proven to be a powerful way to monitor global changes in gene expression patterns. Here, we present an approach that extends the current utility of cDNA arrays to allow the evaluation of the relative roles of transcription and mRNA turnover in governing gene expression on a global basis, compared with current individual gene-by-gene analyses. This method, which involves comparison of large-scale hybridization patterns generated with steady-state mRNA versus newly transcribed (nuclear run-on) RNA, was used to demonstrate the importance of mRNA turnover in regulating gene expression following several conditions of stress.
trans, c, r, i, p, t, ion, stress response, gene expression
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王文恭, Wengong Wang‡, Xiaoling Yang‡, Isabel Lo´pez de Silanes‡, David Carling§¶, and Myriam Gorospe‡〢
THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 278, No.29, Issue of July 18, pp. 27016-27023, 2003,-0001,():
-1年11月30日
Cytoplasmic export of the RNA-binding protein HuR, a process that critically regulates its function, was recently shown to be inhibited by the AMP-activated protein kinase (AMPK). In the present investigation, treatment of human fibroblasts with AMPK activators such as 5-amino-imidazole-4-carboxamide riboside, antimycin A, and sodium azide inhibited cell growth and lowered the expression of proliferative genes. As anticipated, AMPK activation also decreased both the cytoplasmic HuR levels and the association of HuR with target radiolabeled transcripts encoding such proliferative genes. HuR function was previously shown to be implicated in the maintenance of a "young cell" phenotype in models of replicative cellular senescence. We therefore postulated that AMPK activation in human fibroblasts might contribute to the implementation of the senescence phenotype through mechanisms that included a reduction in HuR cytoplasmic presence. Indeed, AMP:ATP ratios were 2-3-fold higher in senescent fibroblasts compared with young fibroblasts. Accordingly, in vitro senescence was accompanied by a marked elevation in AMPK activity. Evidence that increased AMPK activity directly contributed to the implementation of the senescent phenotype was obtained through two experimental approaches. First, use of AMPK activators triggered senescence characteristics in ibroblasts, such as the acquisition of senescence-associated β-galactosidase (β-gal) activity and increased p16INK4a expression. Second, infection of cells with an adenoviral vector that expresses active AMPK increased senescence-associated β-gal activity, whereas infection with an adenovirus that expresses dominant-negative AMPK decreased senescence-associated β-gal activity. Together, our results indicate that AMPK activation can cause premature fibroblast senescence through mechanisms that likely involve reduced HuR function.
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王文恭, Stefanie Galbán, Jennifer L. Martindale, Krystyna Mazan-Mamczarz, Isabel López de Silanes, Jinshui Fan, Wengong Wang, Jochen Decker, and Myriam Gorospe *
MOLECULAR AND CELLULAR BIOLOGY, Oct. 2003, p. 7083-7095,-0001,():
-1年11月30日
A recent analysis of gene expression in renal cell carcinoma cells led to the identification of mRNAs whose translation was dependent on the presence of the von Hippel-Lindau (VHL) tumor suppressor gene product, pVHL. Here, we investigate the finding that pVHL-expressing RCC cells (VHL+) exhibited elevated levels of polysome-associated p53 mRNA and increased p53 protein levels compared with VHL-defective (VHL-) cells. Our findings indicate that p53 translation is specifically heightened in VHL+ cells, given that (i) p53 mRNA abundance in VHL+ and VHL- cells was comparable, (ii) p53 degradation did not significantly influence p53 expression, and (iii) p53 synthesis was markedly induced in VHL+ cells. Electrophoretic mobility shift and immunoprecipitation assays to detect endogenous and radiolabeled p53 transcripts revealed that the RNAbinding protein HuR, previously shown to regulate mRNA turnover and translation, was capable of binding to the 3' untranslated region of the p53 mRNA in a VHL-dependent fashion. Interestingly, while whole-cell levels of HuR in VHL+ and VHL- cells were comparable, HuR was markedly more abundant in the cytoplasmic and polysome-associated fractions of VHL+ cells. In keeping with earlier reports, the elevated cytoplasmic HuR in VHL+ cells was likely due to the reduced AMP-activated kinase activity in these cells. Demonstration that HuR indeed contributed to the increased expression of p53 in VHL+ cells was obtained through use of RNA interference, which effectively reduced HuR expression and in turn caused marked decreases in p53 translation and p53 abundance. Taken together, our findings support a role for pVHL in elevating p53 expression, implicate HuR in enhancing VHL-mediated p53 translation, and suggest that VHL-mediated p53 upregulation may contribute to pVHL's tumor suppressive functions in renal cell carcinoma.
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