Interactions of the F1-ATPase subunits from Escherichia coli detected by the yeast two-hybrid system.
Interactions of the F1-ATPase subunits from Escherichia coli detected by the yeast two-hybrid system.
复制标题
酵母双杂交系统检测到大肠杆菌 F1-ATP 酶亚基的相互作用。
DOI：
10.1016/0005-2728(96)00013-8
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
H. Kanazawa
中科院分区：
文献类型：
--
作者：
C. Moritani;K. Sawada;K. Takemoto;Y. Shin;S. Nemoto;T. Noumi;H. Kanazawa
利用酵母双杂交系统研究了F1-ATPase亚基间的相互作用。将编码大肠杆菌H ~+-ATP酶α、β、γ、δ和γ亚基的基因与酵母GAL 4基因的DNA结合或激活结构域融合，以不同的成对组合导入酵母，并检测编码β-半乳糖苷酶的报告基因的表达。α和β亚基基因的组合以及α和γ亚基基因的组合显示高水平的报告基因表达，而α和δ、β和δ、γ和δ以及δ和γ亚基基因的组合显示弱但显著的报告基因表达。然而，α和γ、β和γ、α和γ β以及β和γ β的组合不诱导报告基因表达。没有一个融合基因单独诱导报告基因表达。这些结果表明，在酵母双杂交系统中，α和β、γ和δ亚基之间存在特异的强相互作用，α和δ、β和δ、γ和δ亚基之间存在弱相互作用。先前鉴定的具有Leu-40至Pro的突变β亚基的影响。分析了Glu-41 → Lys或Pro-332 → Gln取代引起F1-ATPase分子组装缺陷的α-β相互作用。在此系统中，使用Pro-332突变为Gln的β亚基，未观察到α和β亚基之间的相互作用。然而，对于其他两个突变，观察到α-β相互作用。该系统可用于分离F1-ATP酶亚基相互作用缺陷的突变体。
Subunit interactions among the F1-ATPase subunits were studied by the yeast two-hybrid system. Various pairwise combinations of genes encoding α, β, γ, δ and ϵ subunits of Escherichia coli H+-ATPase fused to the DNA-binding or activation domain of the yeast GAL4 gene were introduced into yeast and expression of a reporter gene encoding β-galactosidase was detected. Combinations of the α and β subunit genes, and of the ϵ and γ subunit genes showed high levels of reporter gene expression, while those of α and δ, β and δ, γ and δ, and δ and ϵ demonstrated weak but significant reporter gene expression. However, combinations of α and γ, β and γ, α and ϵ, and β and ϵ did not induce reporter gene expression. None of the fused genes alone induced reporter gene expression. These results suggested that specific and strong interactions between the α and β, γ and ϵ, and weak interactions between the α and δ, β and δ, and γ and δ subunits occurred in yeast cells in the two-hybrid system. Effects of previously identified mutant β subunits with Leu-40 to Pro. Glu-41 to Lys or Pro-332 to Gln substitutions which caused defects in molecular assembly of F1-ATPase were analyzed with regard to α-β interaction. No interaction of the α and β subunits was observed in this system using the β subunit with mutation of Pro-332 to Gln. However, for the other two mutations, α-β interactions were observed. This system may be useful for isolating mutants which have defects in interaction of F1-ATPase subunits.
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DOI：
--
发表时间：
1993
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Pedersen,PL;Amzel,LM
通讯作者：
Amzel,LM
大肠杆菌 F1 ATP 酶的分离的 γ 亚基与 ε 亚基结合。
DOI：
--
发表时间：
1982
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Dunn,SD
通讯作者：
Dunn,SD
大肠杆菌 F1F0-ATP 合酶的交联和标记揭示了 F0 靠近 F1 催化亚基的紧凑亲水部分。
DOI：
--
发表时间：
1983
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Aris,JP;Simoni,RD
通讯作者：
Simoni,RD
大肠杆菌 H-ATP 酶 F0 区 psi 亚基的拓扑、组织和功能。
DOI：
--
发表时间：
1983
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Hermolin,J;Gallant,J;Fillingame,RH
通讯作者：
Fillingame,RH