Developmental Genetics of the Xenopus laevis src Genes

这项拟议的研究的目的是了解pp60src如何调节非洲爪哇卵母细胞的结构和生化特征。我们将研究pp60src基因在卵母细胞中表达时所引起的细胞骨架改变的本质。当黄体酮处理使卵母细胞重新进入细胞周期时，产生激酶构成的pp60src的卵母细胞中酪氨酸磷酸化的模式被改变。通过显微注射周期蛋白mRNA激活卵母细胞的内源性成熟促进因子(MPF)将被用来检验MPF是否是导致底物模式的这种变化的原因。免疫化学和生化方法将被用来确定pp60src分布的变化或修饰是否伴随着底物模式的变化。定点突变将被用来试图确定pp60src中介导孕酮反应的部分。

The goal of the proposed research is to understand how pp60src functions to regulate structural and biochemical features of the Xenopus oocyte. The nature of cytoskeletal changes caused by expression in oocytes of pp60src with constitutive kinase activity will be examined. The pattern of tyrosine phosphorylation in oocytes producing kinase-constitutive pp60src is altered when such oocytes are made to reenter the cell cycle by progesterone treatment. Activation of the oocyte's endogenous maturation promoting factor (MPF) by microinjection of cyclin mRNA will be used to examine whether MPF is responsible for this change in substrate pattern. Immunochemical and biochemical methods will be used to determine whether changes in pp60src distribution or modification accompany the change in substrate pattern. Site directed mutagenesis will be used in an attempt to define the portion of pp60src which mediates the response to progesterone.