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J. Virol., Feb 1995, 756-763, Vol 69, No. 2
M Nacht, SI Reed and JC Alwine
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.
Copyright © 1995, American Society for Microbiology
Simian virus 40 large T antigen affects the Saccharomyces cerevisiae cell cycle and interacts with p34CDC28
Department of Microbiology, University of Pennsylvania, School of Medicine, Philadelphia 19104-6142.
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