Goh Wei Chun, Manel Nicolas, Emerman Michael
Division of Human Biology, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
Virology. 2004 Jan 5;318(1):337-49. doi: 10.1016/j.virol.2003.10.007.
The human immunodeficiency virus (HIV) encodes a gene product, Vpr, which causes infected cells to arrest or delay in the G2 phase of the cell cycle. The arrest in G2 is characterized by low levels of Cyclin B1-p34Cdc2 activity and corresponding inhibitory phosphorylation of p34Cdc2. We find that Vpr directly inhibits the in vitro activity of a phosphatase, Cdc25C, which normally activates Cyclin B1-p34Cdc2. Vpr binds to Cdc25C both in vitro and in mammalian cells. Vpr is also able to inhibit Cdc25C phosphatase activity in vitro although it binds to a site on Cdc25C that is distinct from the catalytic site of the enzyme. Expression of both a catalytically active mutant of Cdc25C that has reduced binding to Vpr as well as a catalytically inactive mutant of Cdc25C that retains binding to Vpr is able to largely overcome Vpr-mediated G2 arrest. Finally, depletion of Cdc25C from cells also renders them partially resistant to the effects of Vpr.
人类免疫缺陷病毒(HIV)编码一种基因产物Vpr,它可使受感染细胞在细胞周期的G2期停滞或延迟。G2期停滞的特征是细胞周期蛋白B1-p34Cdc2活性水平较低以及p34Cdc2相应的抑制性磷酸化。我们发现Vpr直接抑制一种磷酸酶Cdc25C的体外活性,该磷酸酶通常激活细胞周期蛋白B1-p34Cdc2。Vpr在体外和哺乳动物细胞中均与Cdc25C结合。Vpr虽然与Cdc25C上一个不同于该酶催化位点的位点结合,但也能够在体外抑制Cdc25C磷酸酶活性。与Vpr结合减少的Cdc25C催化活性突变体以及保留与Vpr结合的Cdc25C催化无活性突变体的表达,都能够在很大程度上克服Vpr介导的G2期停滞。最后,从细胞中去除Cdc25C也使它们对Vpr的作用产生部分抗性。