Reverte Carlos G, Yuan Lei, Keady Brian T, Lacza Charlemagne, Attfield Kathleen R, Mahon Gwendolyn M, Freeman Benjamin, Whitehead Ian P, Hake Laura E
Biology Department, Boston College, Chestnut Hill, MA 02467, USA.
Dev Biol. 2003 Mar 15;255(2):383-98. doi: 10.1016/s0012-1606(02)00089-1.
XGef was isolated in a screen for proteins interacting with CPEB, a regulator of mRNA translation in early Xenopus development. XGef is a Rho-family guanine nucleotide exchange factor and activates Cdc42 in mammalian cells. Endogenous XGef (58 kDa) interacts with recombinant CPEB, and recombinant XGef interacts with endogenous CPEB in Xenopus oocytes. Injection of XGef antibodies into stage VI Xenopus oocytes blocks progesterone-induced oocyte maturation and prevents the polyadenylation and translation of c-mos mRNA; injection of XGef rescues these events. Overexpression of XGef in oocytes accelerates progesterone-induced oocyte maturation and the polyadenylation and translation of c-mos mRNA. Overexpression of a nucleotide exchange deficient version of XGef, which retains the ability to interact with CPEB, no longer accelerates oocyte maturation or Mos synthesis, suggesting that XGef exchange factor activity is required for the influence of overexpressed XGef on oocyte maturation. XGef overexpression continues to accelerate c-mos polyadenylation in the absence of Mos protein, but does not stimulate MAPK phosphorylation, MPF activation, or oocyte maturation, indicating that XGef may function through the Mos pathway to influence oocyte maturation. These results suggest that XGef may be an early acting component of the progesterone-induced oocyte maturation pathway.
XGef是在一项筛选与CPEB相互作用的蛋白质的实验中分离得到的,CPEB是非洲爪蟾早期发育中mRNA翻译的调节因子。XGef是一种Rho家族鸟嘌呤核苷酸交换因子,在哺乳动物细胞中激活Cdc42。内源性XGef(58 kDa)与重组CPEB相互作用,重组XGef与非洲爪蟾卵母细胞中的内源性CPEB相互作用。将XGef抗体注射到VI期非洲爪蟾卵母细胞中可阻断孕酮诱导的卵母细胞成熟,并阻止c-mos mRNA的多聚腺苷酸化和翻译;注射XGef可挽救这些事件。在卵母细胞中过表达XGef可加速孕酮诱导的卵母细胞成熟以及c-mos mRNA的多聚腺苷酸化和翻译。过表达一种核苷酸交换缺陷型的XGef,其保留了与CPEB相互作用的能力,但不再加速卵母细胞成熟或Mos合成,这表明XGef交换因子活性是过表达的XGef影响卵母细胞成熟所必需的。在没有Mos蛋白的情况下,XGef过表达继续加速c-mos多聚腺苷酸化,但不刺激MAPK磷酸化、MPF激活或卵母细胞成熟,这表明XGef可能通过Mos途径发挥作用来影响卵母细胞成熟。这些结果表明XGef可能是孕酮诱导的卵母细胞成熟途径中的一个早期起作用的成分。