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编码腺酸环酶的S. cerevisiae基因的DNA序列和特征

T Kataoka, D Broek, M Wigler

|December 1, 1985

概括

研究人员克隆了酵母腺酸环酶基因 (CYR1),识别了其催化域和重复的氨基酸序列,与人类糖蛋白相同. 这项工作揭示了酵母细胞信号通路.

科学领域:

分子生物学分子生物学
酵母遗传学酵母遗传学
蛋白质化学蛋白质化学

背景情况:

腺酸环酶是细胞信号通路中的关键酶,调节循环AMP (cAMP) 水平.
编码S. cerevisiae腺酸环酶的基因,CYR1,以前没有被表征.
RAS基因在酵母细胞的生长和活力中起着至关重要的作用.

研究的目的:

为了克隆和描述编码腺酸环酶的S. cerevisiae CYR1基因.
为了确定腺酸环酶蛋白的功能域.
调查酵母酵母中腺酸环酶和RAS基因功能之间的关系.

主要方法:

基因克隆和CYR1.1的基因测序.
在酵母中进行基因破坏实验.
对腺酸环酶活性进行生物化学分析.
酵母腺酸环酶在大肠杆菌中的表达.

主要成果:

CYR1基因编码了一种2026氨基酸蛋白质,其中心重复区域与人血清葡萄糖蛋白相同.
乙酸环酸酶的催化域位于400氨基酸的炭基末端.
过度表达的腺酸环酶拯救了由酵母中RAS基因损失引起的致死性.
在大肠杆菌中产生的重组酵母腺酸环酶并未被RAS蛋白激活.

结论:

CYR1基因编码了S. cerevisiae中唯一的腺酸环酶.
酵母腺酸环酶的结构和功能领域已经被阐明.
在酵母中的腺酸环酶和RAS信号之间存在功能联系,尽管在体外没有观察到RAS蛋白直接激活.

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