Selection of apoptosis-deficient adenovirus E4orf4 mutants in Saccharomyces cerevisiae.
Adenovirus E4orf4 protein has been shown to induce p53-independent, protein phosphatase 2A (PP2A)-dependent apoptosis in transformed cells. Furthermore, E4orf4 also induces toxicity in Saccharomyces cerevisiae in a PP2A-dependent manner (D. Kornitzer and T. Kleinberger, submitted for publication). In this work, we utilized yeast cells to select for nonapoptotic E4orf4 mutants which, in turn, were shown to possess a diminished ability to bind PP2A. The success of this selection system will provide additional apoptosis-relevant mutants for E4orf4 research and strongly supports the relevance of E4orf4-induced toxicity in S. cerevisiae to E4orf4-induced apoptosis in mammalian cells.
Afifi
R
R
Gonda Center of Molecular Microbiology, Bruce Rappaport Faculty of Medicine, Technion, Haifa 31096, Israel.
Sharf
R
R
Shtrichman
R
R
Kleinberger
T
T
D000260:Adenoviruses, Human; D000595:Amino Acid Sequence; D017209:Apoptosis; D002461:Cell Line, Transformed; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D054648:Protein Phosphatase 2; D012441:Saccharomyces cerevisiae; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*
C121517:E4orf4 protein, adenovirus; D014764:Viral Proteins; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2