Mapping the cAMP receptor protein contact site on the α subunit of Escherichia coli RNA polymerase
Mapping the cAMP receptor protein contact site on the α subunit of Escherichia coli RNA polymerase
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绘制大肠杆菌 RNA 聚合酶 α 亚基上的 cAMP 受体蛋白接触位点
DOI：
10.1111/j.1365-2958.1992.tb01437.x
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发表时间：
1992
影响因子：
3.6
通讯作者：
A. Ishihama
中科院分区：
文献类型：
--
作者：
C. Zou;N. Fujita;K. Igarashi;A. Ishihama
大肠杆菌RNA聚合酶α亚基的C末端区域（氨基酸残基236-329）携带正转录因子的接触位点I。为了详细定位cAMP受体蛋白（CRP）的接触位点，我们通过聚合酶链反应（PCR）诱变制备了一个突变型rpoA文库，使每个突变型平均仅在rpoA基因的C末端携带一个突变，然后筛选该文库中lacZ基因表达降低的突变体。在cAMP-CRP存在下，含有突变体a亚基的重组全酶在体外转录galP 1，但不转录lacP 1。几个“Lac‐”突变rpoA基因的DNA序列测定显示，所有突变都聚集在α C末端附近氨基酸残基265和270之间的短片段内。接触位点簇似乎存在于接触位点I区域内，每个包含约5个氨基酸，并在分子通讯中与不同的转录因子应答。
The C‐terminal region (amino acid residues 236–329) of the Escherichia coli RNA polymerase α subunit carries the contact site I for positive transcription factors. For detailed mapping of the contact site for the cAMP receptor protein (CRP), we made a library of mutant rpoA by polymerase chain reaction (PCR) mutagenesis, such that each should carry a single mutation on average and exclusively in the C‐terminal half of the rpoA gene, and then screened this library for mutants with decreased expression of the lacZ gene. Reconstituted holoenzyme containing the mutant a subunits transcribed galP1 but not lacP1 in vitro in the presence of cAMP–CRP. DNA sequence determination of several ‘Lac‐’ mutant rpoA genes revealed that all had mutations clustered within a short segment near the C‐terminus of α between amino acid residues 265 and 270. A cluster of contact sites appear to exist within the contact site I region, each comprising of about five amino acids and responding in molecular communication with a different transcription factor(s).
单克隆抗体作为大肠杆菌 RNA 聚合酶 α 亚基拓扑排列的探针。
影响因子：
2.9
作者：
Riftina,F;DeFalco,E;Krakow,JS
通讯作者：
Krakow,JS
抗 α 单克隆抗体对大肠杆菌 RNA 聚合酶与 lac 启动子相互作用的影响。
影响因子：
2.9
作者：
Riftina,F;DeFalco,E;Krakow,JS
通讯作者：
Krakow,JS