The overall aim of this research is the determination of the functions of SV40 small-t antigen and of two cellular proteins with which it associates. Major effort will be placed on investigations of the metabolic patterns of cells that express small-t antigen, because recent studies have shown that antimitochondrial drugs and ionophores modify growth patterns observed in CV-1 cells which relate to the presence of small-t antigen. Plasma membrane functions, such as transport and membrane potential, will also be examined and related to small-t antigen. We will also pursue the observation that one of the cellular proteins, 32K, appears to be binding S-adenosylmethionine (SAM). This raises the possibility that the 32K protein may be an enzyme which uses SAM as a substrate, and may provide a major clue concerning the function of at least one of the cellular proteins and, indirectly, the function of small-t antigen. Purifications of small-t antigen and the cellular proteins will be continued and scaled-up, with particular emphasis on purification of the soluble small-t antigen expressed by a bacterial clone. Isolation of small-t antigen which retains the ability to interact with the cellular proteins is a priority. Additional efforts include development of hybridoma cell lines that produce antibodies that recognize the small-t antigen cellular protein complex efficiently, or that recognize the unique region of small-t antigen. If the latter is obtained, attempts to identify and active protein fragment from small-t antigen in infected cells will be made. Potential function of such a fragment is also being approached using a bacterial clone that contains DNA sequences only from this region of small-t antigen, and from which small-t unique sequences can be expressed in bacteria.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA021327-12
Application #
3165528
Study Section
Experimental Virology Study Section (EVR)
Project Start
1977-05-01
Project End
1992-04-30
Budget Start
1989-05-01
Budget End
1990-04-30
Support Year
12
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Northwestern University at Chicago
Department
Type
School of Medicine & Dentistry
DUNS #
005436803
City
Chicago
State
IL
Country
United States
Zip Code
60611
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Genevaux, Pierre; Lang, Florence; Schwager, Francoise et al. (2003) Simian virus 40 T antigens and J domains: analysis of Hsp40 cochaperone functions in Escherichia coli. J Virol 77:10706-13
Montano, X; Rundell, K (2001) Role of SV40 small t in cell lysis, transformation, and signaling. Methods Mol Biol 165:229-42
Rundell, K; Parakati, R (2001) The role of the SV40 ST antigen in cell growth promotion and transformation. Semin Cancer Biol 11:5-13

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