Biochemical and cytological changes associated with expression of deregulated pp60src in Xenopus oocytes
Biochemical and cytological changes associated with expression of deregulated pp60src in Xenopus oocytes
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与非洲爪蟾卵母细胞中 pp60src 表达失调相关的生化和细胞学变化
DOI：
10.1128/mcb.12.12.5485-5498.1992
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发表时间：
1992
影响因子：
5.3
通讯作者：
R. Steele
中科院分区：
文献类型：
--
作者：
T. F. Unger;R. Steele
我们研究了具有组成性激酶活性的爪蟾pp 60 c-src对非洲爪蟾卵母细胞形态和成熟的影响。当将编码这种失调变体的RNA注射到VI期卵母细胞中时，我们观察到卵母细胞皮质的总体变化。这种改变涉及色素聚集和注射部位近端大面积皮质内陷。在注射了编码野生型pp 60 c-src的RNA的卵母细胞中没有观察到这种现象。我们已经将这种现象与84-和100-kDa蛋白质的酪氨酸磷酸化相关联。这些磷酸化的蛋白质共定位与改变在卵母细胞皮层时，生化和免疫细胞化学方法测定。无论是色素聚集，也没有磷酸化的84-和100-kDa的蛋白质中观察到的卵母细胞表达的非豆蔻酰化版本的失调pp 60 c-src。表达失调的爪蟾fyn，src家族成员，导致表型与失调的src相似。然而，在注射fyn的卵母细胞中，酪氨酸磷酸化的蛋白质比注射src的卵母细胞多得多。孕激素刺激的卵母细胞表达失调pp 60 c-src导致酪氨酸磷酸化蛋白的数量增加。这种变化可能代表了pp 60 src对成熟卵母细胞细胞周期恢复的反应。这些数据表明，卵母细胞可能是一个特别有用的系统，用于研究pp 60 c-src在细胞骨架结构调节和细胞周期相关事件调节中的作用。
We have examined the effects of Xenopus pp60c-src with constitutive kinase activity on the morphology and maturation of Xenopus laevis oocytes. When RNA encoding this deregulated variant was injected into stage VI oocytes, we observed a gross alteration in the cortex of the oocyte. This alteration involved aggregation of pigment and invagination of the cortex in a large area proximal to the site of injection. This phenomenon was not seen in oocytes injected with RNA encoding wild-type pp60c-src. We have correlated this phenomenon with the tyrosine phosphorylation of 84- and 100-kDa proteins. These phosphorylated proteins colocalized with the alteration in the oocyte cortex when assayed by both biochemical and immunocytochemical methods. Neither the pigment aggregation nor phosphorylation of the 84- and 100-kDa proteins was observed in oocytes expressing a nonmyristoylated version of the deregulated pp60c-src. Expression of deregulated Xenopus fyn, a src-family member, resulted in a phenotype similar to that seen with deregulated src. However, in the fyn-injected oocytes, many more proteins were phosphorylated on tyrosine than in the src-injected oocytes. Progesterone stimulation of oocytes expressing deregulated pp60c-src resulted in an increase in the number of tyrosine-phosphorylated proteins. This change may represent the response of pp60src to the resumption of the cell cycle in maturing oocytes. These data suggest that the oocyte may be a particularly useful system for investigating the role of pp60c-src in the regulation of cytoskeletal structure and in the regulation of events associated with the cell cycle.