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J. Virol., Feb 1995, 756-763, Vol 69, No. 2
Copyright © 1995, American Society for Microbiology

Simian virus 40 large T antigen affects the Saccharomyces cerevisiae cell cycle and interacts with p34CDC28

M Nacht, SI Reed and JC Alwine
Department of Microbiology, University of Pennsylvania, School of Medicine, Philadelphia 19104-6142.

Simian virus 40 tumor (T) antigen, an established viral oncoprotein, causes alterations in cell growth control through interacting with, and altering the function of, cellular proteins. To examine the effects of T antigen on cell growth control, and to identify the cellular proteins with which it may functionally interact, T antigen was expressed in the budding yeast Saccharomyces cerevisiae. The yeast cells expressing T antigen showed morphological alterations as well as growth inhibition attributable, at least in part, to a lag in progression from G1 to S. This point in the cell cycle is also known to be affected by T antigen in mammalian cells. Both p34CDC28 and p34CDC2Hs were shown to bind to a chimeric T antigen-glutathione S-transferase fusion protein, indicating that T antigen interacts directly with cell cycle proteins which control the G1 to S transition. This interaction was confirmed by in vivo cross-linking experiments, in which T antigen and p34CDC28 were coimmunoprecipitated from extracts of T-antigen-expressing yeast cells. These immunoprecipitated complexes could phosphorylate histone H1, indicating that kinase activity was retained. In addition, in autophosphorylation reactions, the complexes phosphorylated a novel 60- kDa protein which appeared to be underphosphorylated (or underrepresented) in p34CDC28-containing complexes from cells which did not express T antigen. These results suggest that T antigen interacts with p34CDC28 and alters the kinase function of p34CDC28-containing complexes. These events correlate with alterations in the yeast cell cycle at the G1 to S transition.


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Copyright © 1995 by the American Society for Microbiology. All rights reserved.