Genetic Selection of Mammalian Adenylyl Cyclases Insensitive to Stimulation by Gsα *

Genetic Selection of Mammalian Adenylyl Cyclases Insensitive to Stimulation by Gsα *
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对 Gsα 刺激不敏感的哺乳动物腺苷酸环化酶的遗传选择 *

DOI:
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发表时间:
1998
影响因子:
4.8
通讯作者:
R. Taussig
R. Taussig
中科院分区:
生物学2区
文献类型:
--
作者:
G. Zimmermann;Dongmei Zhou;R. Taussig

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我们描述了一种遗传系统的发展,该遗传系统允许分离突变的哺乳动物腺苷酸环化酶对G蛋白亚基的反应有缺陷,从而允许鉴定环化酶中负责识别这些调节因子的结构元件。哺乳动物V型腺苷酸环化酶在环化酶缺失酵母菌株中的表达可以有条件地补充该菌株的致死表型。表达V型腺苷酸环化酶的酵母只有在共表达gsa α或在培养基中添加forskolin激活环化酶时才能生长;然而,在两种活化剂存在或在基础条件下观察到生长停滞。利用这个遗传系统,我们分离了25个对gsa α反应有缺陷的腺苷酸环化酶突变体。这些突变体的序列分析和生化特性已经确定了环化酶的两个细胞质区域的残基,这些残基参与了Gsα的特异性结合和调节。
We describe the development of a genetic system allowing for the isolation of mutant mammalian adenylyl cyclases defective in their responses to G protein subunits, thus allowing for the identification of structural elements within the cyclase that are responsible for the recognition of these regulators. Expression of mammalian type V adenylyl cyclase in a cyclase-deleted yeast strain can conditionally complement the lethal phenotype of this strain. Type V adenylyl cyclase-expressing yeast grow only when the cyclase is activated by coexpression of Gsα or addition of forskolin to the medium; however, growth arrest is observed in the presence of both activators or under basal conditions. Utilizing this genetic system, we have isolated 25 adenylyl cyclase mutants defective in their response to Gsα. Sequence analysis and biochemical characterization of these mutants have identified residues in both cytoplasmic domains of the cyclase that are involved in the specific binding of and regulation by Gsα.
DOI: 10.1126/science.1962211
发表时间: 1991-12
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影响因子: 56.9
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DOI: --
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DOI: --
发表时间: 1993
期刊: The Journal of biological chemistry
影响因子: --
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