Simian Virus 40 Large T Antigen Binds a Novel Bcl-2 Homology Domain 3-containing Proapoptosis Protein in the Cytoplasm*

A 193-kDa SV40 large T antigen (T-Ag)-binding protein, designated p193, was identified and cloned. Inspection of the deduced amino acid sequence revealed the presence of a short motif similar to the Bcl-2 homology (BH) domain 3, suggesting that p193 may be a member of a family of apoptosis promoting proteins containing only BH3 motifs. In support of this, p193 expression promoted apoptosis in NIH-3T3 cells. Deletion of the BH3 motif abolished p193 apoptosis activity. p193-induced apoptosis was antagonized by co-expression of Bcl-XL. Immune cytologic analysis indicated that p193 is localized to the cytoplasm of transfected cells. p193-induced apoptosis was also antagonized by co-expression of T-Ag, which resulted in the cytoplasmic localization of both proteins. The p193 binding site was mapped to an N-terminal region of T-Ag previously implicated in transforming activity. These results suggest that T-Ag possesses an antiapoptosis activity, independent of p53 sequestration, which is actuated by T-Ag/p193 binding in the cytoplasm.

[1]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[2]  J. Ross,et al.  Loss of a gp130 Cardiac Muscle Cell Survival Pathway Is a Critical Event in the Onset of Heart Failure during Biomechanical Stress , 1999, Cell.

[3]  S. Vatner,et al.  Apoptosis of Cardiac Myocytes in Gsα Transgenic Mice , 1999 .

[4]  Y. Lazebnik,et al.  Caspases: enemies within. , 1998, Science.

[5]  V. Dixit,et al.  Death receptors: signaling and modulation. , 1998, Science.

[6]  S. Cory,et al.  The Bcl-2 protein family: arbiters of cell survival. , 1998, Science.

[7]  M. Soonpaa,et al.  Survey of studies examining mammalian cardiomyocyte DNA synthesis. , 1998, Circulation research.

[8]  P. Anversa,et al.  Myocyte death and growth in the failing heart. , 1998, Laboratory investigation; a journal of technical methods and pathology.

[9]  S. Izumo,et al.  Apoptosis: basic mechanisms and implications for cardiovascular disease. , 1998, Circulation research.

[10]  R. Lang,et al.  Dilated cardiomyopathy in transgenic mice expressing a dominant-negative CREB transcription factor in the heart. , 1998, The Journal of clinical investigation.

[11]  H. Horvitz,et al.  The C. elegans Protein EGL-1 Is Required for Programmed Cell Death and Interacts with the Bcl-2–like Protein CED-9 , 1998, Cell.

[12]  Charles A. Thomas,et al.  The human oncoprotein MDM2 arrests the cell cycle: elimination of its cell‐cycle‐inhibitory function induces tumorigenesis , 1998, The EMBO journal.

[13]  R. Peshock,et al.  Cardiac failure in transgenic mice with myocardial expression of tumor necrosis factor-alpha. , 1998, Circulation.

[14]  S. Srinivasula,et al.  Blk, a BH3-containing Mouse Protein That Interacts with Bcl-2 and Bcl-xL, Is a Potent Death Agonist* , 1998, The Journal of Biological Chemistry.

[15]  A. Strasser,et al.  Bim: a novel member of the Bcl‐2 family that promotes apoptosis , 1998, The EMBO journal.

[16]  L. Truong,et al.  Adenoviral delivery of E2F-1 directs cell cycle reentry and p53-independent apoptosis in postmitotic adult myocardium in vivo. , 1997, The Journal of clinical investigation.

[17]  A. Koretsky,et al.  Dilated Cardiomyopathy in Transgenic Mice With Cardiac-Specific Overexpression of Tumor Necrosis Factor-α , 1997 .

[18]  K. Webster,et al.  Adenovirus E1A Inhibits Cardiac Myocyte-specific Gene Expression through Its Amino Terminus* , 1997, The Journal of Biological Chemistry.

[19]  I. M. Marks,et al.  The amino-terminal transforming region of simian virus 40 large T and small t antigens functions as a J domain , 1997, Molecular and cellular biology.

[20]  C. Cole,et al.  Identification of a novel antiapoptotic functional domain in simian virus 40 large T antigen , 1997, Journal of virology.

[21]  P. Silver,et al.  DnaJ/hsp40 chaperone domain of SV40 large T antigen promotes efficient viral DNA replication. , 1997, Genes & development.

[22]  Y. Hsu,et al.  Cytosol-to-membrane redistribution of Bax and Bcl-X(L) during apoptosis. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[23]  N. Inohara,et al.  harakiri, a novel regulator of cell death, encodes a protein that activates apoptosis and interacts selectively with survival‐promoting proteins Bcl‐2 and Bcl‐XL , 1997, The EMBO journal.

[24]  W. Henzel,et al.  Mouse RAD50 Has Limited Epitopic Homology to p53 and Is Expressed in the Adult Myocardium* , 1996, The Journal of Biological Chemistry.

[25]  C. Milliman,et al.  BID: a novel BH3 domain-only death agonist. , 1996, Genes & development.

[26]  M. Abdellatif,et al.  Human E2F-1 reactivates cell cycle progression in ventricular myocytes and represses cardiac gene transcription. , 1996, Developmental biology.

[27]  Y. Kaneda,et al.  Fusigenic liposome-mediated DNA transfer into cardiac myocytes. , 1996, Journal of molecular and cellular cardiology.

[28]  G. Koh,et al.  Genetically selected cardiomyocytes from differentiating embronic stem cells form stable intracardiac grafts. , 1996, The Journal of clinical investigation.

[29]  C. Prives,et al.  p53: puzzle and paradigm. , 1996, Genes & development.

[30]  R. Kitsis,et al.  Induction of DNA synthesis and apoptosis in cardiac myocytes by E1A oncoprotein , 1996, The Journal of cell biology.

[31]  A. Shevchenko,et al.  Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. , 1996, Analytical chemistry.

[32]  E. White,et al.  The E1B 19K protein blocks apoptosis by interacting with and inhibiting the p53-inducible and death-promoting Bax protein. , 1996, Genes & development.

[33]  N. L. Thangue,et al.  E2F and the molecular mechanisms of early cell-cycle control. , 1996 .

[34]  Andreas Radbruch,et al.  Isolation of full-size mRNA from ethanol-fixed cells after cellular immunofluorescence staining and fluorescence-activated cell sorting (FACS). , 1995, Cytometry.

[35]  J. M. Boyd,et al.  Bik, a novel death-inducing protein shares a distinct sequence motif with Bcl-2 family proteins and interacts with viral and cellular survival-promoting proteins. , 1995, Oncogene.

[36]  M. Soonpaa,et al.  Developmental expression of p107 mRNA and evidence for alternative splicing of the p107 (RBL1) gene product. , 1995, Genomics.

[37]  J. Martinou,et al.  Cloning of a bcl-2 homologue by interaction with adenovirus E1B 19K , 1995, Nature.

[38]  Matthew J. Brauer,et al.  Modulation of apoptosis by the widely distributed Bcl-2 homologue Bak , 1995, Nature.

[39]  G. Evan,et al.  Induction of apoptosis by the Bcl-2 homologue Bak , 1995, Nature.

[40]  L. Kirshenbaum,et al.  Adenovirus E1A Represses Cardiac Gene Transcription and Reactivates DNA Synthesis in Ventricular Myocytes, via Alternative Pocket Protein- and p300-binding Domains (*) , 1995, The Journal of Biological Chemistry.

[41]  G. Skuse,et al.  Viral oncoprotein binding to pRB, p107, p130, and p300. , 1995, Virus research.

[42]  S. Korsmeyer,et al.  Bad, a heterodimeric partner for Bcl-xL and Bcl-2, displaces bax and promotes cell death , 1995, Cell.

[43]  J. M. Boyd,et al.  Adenovirus E1B 19 kDa and Bcl-2 proteins interact with a common set of cellular proteins , 1994, Cell.

[44]  G. Koh,et al.  Tumor suppressor gene expression during normal and pathologic myocardial growth. , 1994, The Journal of biological chemistry.

[45]  J. Stults,et al.  Analysis of Two-Dimensional Gel Proteins by Mass Spectrometry and Microsequencing , 1994 .

[46]  J. D. Thompson,et al.  Evidence for a protein domain superfamily shared by the cyclins, TFIIB and RB/p107. , 1994, Nucleic acids research.

[47]  D. Kirsch,et al.  Sequenching of peptides in a time-of-flight mass spectrometer: evaluation of postsource decay following matrix-assisted laser desorption ionisation (MALDI) , 1994 .

[48]  A. Duncan,et al.  The adenovirus E1A-associated 130-kD protein is encoded by a member of the retinoblastoma gene family and physically interacts with cyclins A and E. , 1993, Genes & development.

[49]  G. Hannon,et al.  Isolation of the Rb-related p130 through its interaction with CDK2 and cyclins. , 1993, Genes & development.

[50]  S. Korsmeyer,et al.  Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programed cell death , 1993, Cell.

[51]  E. Moran.  Interaction of adenoviral proteins with pRB and p53 , 1993, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[52]  M. J. Tevethia,et al.  Association of p53 binding and immortalization of primary C57BL/6 mouse embryo fibroblasts by using simian virus 40 T-antigen mutants bearing internal overlapping deletion mutations , 1993, Journal of virology.

[53]  J. Nevins,et al.  E2F: a link between the Rb tumor suppressor protein and viral oncoproteins. , 1992, Science.

[54]  C. Cole,et al.  Transformation of a continuous rat embryo fibroblast cell line requires three separate domains of simian virus 40 large T antigen , 1992, Journal of virology.

[55]  M. Imperiale,et al.  Simian virus 40 large T antigen stably complexes with a 185-kilodalton host protein , 1992, Journal of virology.

[56]  J. Sambrook,et al.  Protein folding in the cell , 1992, Nature.

[57]  M. Ewen,et al.  Molecular cloning, chromosomal mapping, and expression of the cDNA for p107, a retinoblastoma gene product-related protein , 1991, Cell.

[58]  K. Chien,et al.  Terminally differentiated neonatal rat myocardial cells proliferate and maintain specific differentiated functions following expression of SV40 large T antigen. , 1988, The Journal of biological chemistry.

[59]  Wen-Hwa Lee,et al.  SV40 large tumor antigen forms a specific complex with the product of the retinoblastoma susceptibility gene , 1988, Cell.

[60]  L. Field,et al.  Atrial natriuretic factor-SV40 T antigen transgenes produce tumors and cardiac arrhythmias in mice. , 1988, Science.

[61]  J. Sklar,et al.  Cloning and structural analysis of cDNAs for bcl-2 and a hybrid bcl-2/immunoglobulin transcript resulting from the t(14;18) translocation , 1986, Cell.

[62]  D. Wynford‐Thomas,et al.  Heterogeneity in distribution and content of p53 in SV40-transformed mouse fibroblasts. , 1986, Experimental cell research.

[63]  S. Korsmeyer,et al.  Cloning the chromosomal breakpoint of t(14;18) human lymphomas: clustering around Jh on chromosome 14 and near a transcriptional unit on 18 , 1985, Cell.

[64]  P. Nowell,et al.  Cloning of the chromosome breakpoint of neoplastic B cells with the t(14;18) chromosome translocation. , 1984, Science.

[65]  N. Ringertz,et al.  Expression of sv40 t antigen during the cell cycle of SV40‐transformed cells , 1975, International journal of cancer.

[66]  K. Vousden,et al.  Mechanisms of p53-mediated apoptosis , 1999, Cellular and Molecular Life Sciences CMLS.

[67]  R. Weinberg,et al.  The role of RB in cell cycle control. , 1995, Progress in cell cycle research.

[68]  C. Watanabe,et al.  Computer analysis of automated Edman degradation and amino acid analysis data. , 1987, Journal of chromatography.