SV40 large T antigen (T Ag) is capable of transforming a variety of normal cells into cancer cells. Cellular transformation is dependent on T Ag's ability to interact with key cellular regulatory proteins including pRb and p53. This laboratory has recently cloned p185, a novel T Ag associated protein. Furthermore, we have identified a small region of T Ag that is required for binding to p185. Mutation of this region of T Ag eliminates p185 binding but does not affect T Ag's ability to bind and inactivate pRb and p53. T Ag mutants that fail to bind to p185 are unable to transform primary mouse embryo fibroblasts suggesting that T Ag binding to p185 is required for transformation. p185 has significant homology to the cullin family of E3 ubiquitin ligases and to Apc10/Doc1, a component of the anaphase-promoting complex (APC). Cullins and the APC promote the degradation of specific protein substrates by serving as a scaffold for protein substrates and E2 ubiquitin conjugating enzymes. Preliminary Studies supports the hypothesis that p185 is a novel E3 ubiquitin ligase. In addition to cloning p185, we have cloned two p185-associated proteins including p281, highly related to p185, and an F-box containing protein, Fbw6/Fbx29. These results suggest that T Ag binds specifically to a novel cullin with potential to act as an E3 ubiquitin ligase and that this interaction is critical for cellular transformation. This proposal seeks to determine how p185 contributes to T Ag-mediated transformation. In addition, this proposal seeks to identify additional binding proteins of p185, to characterize the p185 ubiquitinating activity, and to determine if p185 serves as a tumor suppressor in a knockout mouse model.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
1R01CA093804-01A1
Application #
6542212
Study Section
Experimental Virology Study Section (EVR)
Program Officer
Blair, Donald G
Project Start
2002-07-01
Project End
2007-06-30
Budget Start
2002-07-01
Budget End
2003-06-30
Support Year
1
Fiscal Year
2002
Total Cost
$368,068
Indirect Cost
Name
Dana-Farber Cancer Institute
Department
Type
DUNS #
149617367
City
Boston
State
MA
Country
United States
Zip Code
02215
Berrios, Christian; Jung, Joonil; Primi, Blake et al. (2015) Malawi polyomavirus is a prevalent human virus that interacts with known tumor suppressors. J Virol 89:857-62
Cheng, Jingwei; Rozenblatt-Rosen, Orit; Paulson, Kelly G et al. (2013) Merkel cell polyomavirus large T antigen has growth-promoting and inhibitory activities. J Virol 87:6118-26
DeCaprio, James A; Garcea, Robert L (2013) A cornucopia of human polyomaviruses. Nat Rev Microbiol 11:264-76
Duda, David M; Olszewski, Jennifer L; Tron, Adriana E et al. (2012) Structure of a glomulin-RBX1-CUL1 complex: inhibition of a RING E3 ligase through masking of its E2-binding surface. Mol Cell 47:371-82
Sadasivam, Subhashini; Duan, Shenghua; DeCaprio, James A (2012) The MuvB complex sequentially recruits B-Myb and FoxM1 to promote mitotic gene expression. Genes Dev 26:474-89
Rozenblatt-Rosen, Orit; Deo, Rahul C; Padi, Megha et al. (2012) Interpreting cancer genomes using systematic host network perturbations by tumour virus proteins. Nature 487:491-5
Tron, Adriana E; Arai, Takehiro; Duda, David M et al. (2012) The glomuvenous malformation protein Glomulin binds Rbx1 and regulates cullin RING ligase-mediated turnover of Fbw7. Mol Cell 46:67-78
Fine, Debrah A; Rozenblatt-Rosen, Orit; Padi, Megha et al. (2012) Identification of FAM111A as an SV40 host range restriction and adenovirus helper factor. PLoS Pathog 8:e1002949
Müller, Gerd A; Quaas, Marianne; Schümann, Michael et al. (2012) The CHR promoter element controls cell cycle-dependent gene transcription and binds the DREAM and MMB complexes. Nucleic Acids Res 40:1561-78
Tschöp, Katrin; Conery, Andrew R; Litovchick, Larisa et al. (2011) A kinase shRNA screen links LATS2 and the pRB tumor suppressor. Genes Dev 25:814-30

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