Developmental Genetics of the Xenopus laevis src Genes

本研究的目的是了解pp60src如何调节爪蟾卵母细胞的结构和生化特征。我们将研究具有组成激酶活性的pp60src在卵母细胞中的表达所引起的细胞骨架变化的性质。当孕激素处理使卵母细胞重新进入细胞周期时，产生激酶组成pp60src的卵母细胞中的酪氨酸磷酸化模式发生改变。通过细胞周期蛋白mRNA的微注射激活卵母细胞的内源性成熟促进因子（MPF）将被用来检查MPF是否对底物模式的这种变化负责。免疫化学和生化方法将用于确定pp60src分布或修饰的变化是否伴随底物模式的变化。位点定向诱变将用于确定pp60src中介导对黄体酮反应的部分。

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.