Phosphorylation of the retinoblastoma protein is modulated in mouse kidney cells infected with polyomavirus.
Lytic infection with polyomavirus, an oncogenic DNA-containing virus, leads in G0-arrested primary baby mouse kidney (BMK) cell cultures to a mitotic host reaction. In the present work, we examined the expression of the retinoblastoma gene (RB) and of its product (Rb) in virus-infected BMK with the aim of correlating its modulation with the sequential activation of cellular processes leading to the induction of S phase by virus. In contrast to cell cycle-regulated genes whose expression is induced by viral infection, expression of RB is not altered during the transition from G0/G1 to S phase. In BMK cell cultures irreversibly arrested in the G0 phase of the cell cycle, an unphosphorylated species is the only detectable form of the RB protein (Rb). Time course analysis showed that in polyoma-infected cells induced to re-enter the S phase of the cell cycle the appearance of the phosphorylated forms of Rb coincided in time with the accumulation of large T antigen and preceded DNA synthesis. During the late phase of infection, the majority of Rb was present as phosphorylated forms. Ongoing DNA synthesis was not required for the cells to phosphorylate Rb, indicating that this post-translational modification takes place during the activation of the cellular DNA-synthesizing apparatus. Using hamster anti-polyoma tumor serum, it was observed that the underphosphorylated form of Rb co-precipitated with polyoma large T antigen extracted from infected cells late during infection. Our data add more evidence to the proposal that interactions between viral early proteins encoded by DNA tumor viruses and the product of RB may play a pivotal role in the mitogenic effect induced by viral infection.
Khandjian
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EW
Département de biochimie, Faculté de médecine, Université Laval, Québec, Canada.
Tremblay
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D000818:Animals; D000957:Antigens, Viral, Tumor / Q000502:physiology; D015152:Blotting, Northern; D015153:Blotting, Western; D002455:Cell Division / Q000235:genetics; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D015180:Electrophoresis, Gel, Two-Dimensional; D015967:Gene Expression Regulation, Viral; D051379:Mice; D010766:Phosphorylation; D011120:Polyomavirus; D011499:Protein Processing, Post-Translational; D012313:RNA / Q000032:analysis; D016160:Retinoblastoma Protein / Q000378:metabolism*
D000957:Antigens, Viral, Tumor; D016160:Retinoblastoma Protein; D012313:RNA; D004247:DNA