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对CAP的克隆和表征,S. cerevisiae基因编码了70kd的腺酸环酶相关蛋白质
J Field1, A Vojtek, R Ballester
1Cold Spring Harbor Laboratory, New York 11724.
Cell
|April 20, 1990
概括
循环酶相关蛋白 (CAP) 对于酵母中的RAS响应性腺烯基循环酶信号传递至关重要. 缺乏CAP的突变者也表现出生长缺陷和形态变化,这表明细胞作用更广泛.
科学领域:
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
- 生物化学 生物化学
背景情况:
- 在Saccharomyces cerevisiae中的腺基酶是一种多个子单元的酶.
- RAS/基基环酶通路调节细胞对各种刺激的反应.
研究的目的:
- 克隆和表征酵母腺酸环酶的循环酶相关蛋白 (CAP) 子单元.
- 研究CAP在RAS介导信号传递和其他细胞过程中的作用.
主要方法:
- 使用大肠杆菌中的酵母表达库克隆CAPcDNA.
- 用抗血清对净化CAP进行查.
- 在缺乏CAP的酵母膜中分析腺环酶活性.
- 对CAP突变体的表型分析.
主要成果:
- 隔离了一种编码为526氨基酸CAP蛋白的cDNA;它与已知的序列没有同质性.
- 缺乏CAP的酵母在体外表现出RAS2刺激的腺环酶活性受损.
- 缺乏CAP的突变体表现出温度敏感性,无法在丰富的介质上生长,饥饿敏感性和细胞形态的改变.
结论:
- 在酵母中,CAP是RAS响应性腺环酶信号通路的关键组成部分.
- 除了RAS信号外,CAP还发挥作用,影响生长,形态和应激反应.
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