DNA-binding domain of the RepE initiator protein of mini-F plasmid: involvement of the carboxyl-terminal region
DNA-binding domain of the RepE initiator protein of mini-F plasmid: involvement of the carboxyl-terminal region
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mini-F 质粒 RepE 起始蛋白的 DNA 结合域：羧基末端区域的参与
DOI：
10.1128/jb.177.8.1994-2001.1995
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发表时间：
1995
影响因子：
3.2
通讯作者：
C. Wada
中科院分区：
文献类型：
--
作者：
F. Matsunaga;Y. Kawasaki;M. Ishiai;K. Nishikawa;T. Yura;C. Wada
RepE启动蛋白（251个残基）在大肠杆菌中对mini-F复制至关重要，具有两个主要功能：从ori2开始DNA复制和自我抑制RepE转录。虽然起始是由与ori2内含子结合的RepE单体介导的（直接重复），但自体抑制是由与RepE操作符结合的二聚体介导的，该操作符包含与内含子相关的反向重复序列。我们现在报道，RepE与这些DNA位点的结合主要是由该蛋白的c端区域决定的。突变的RepE蛋白缺乏n -末端33（或更多）残基或c -末端7（或更多）残基，首先显示在与ori2和操作dna的结合中存在缺陷。然而，直接筛选和分析特异性影响与ori2 iteron结合的突变型repe发现，突变（主要是氨基酸取代）仅发生在c端区域（残基168 ~ 242）。这些突变蛋白显示出与ori2的结合减少，与操作者没有可检测到的结合。因此，尽管截断RepE的任何一端都可以破坏dna结合活性，但c端区域似乎代表了RepE的主要dna结合域，对于ori2和操作符来说都是如此。类似的dna结合域似乎在某些相关质粒的启动蛋白中是保守的。
The RepE initiator protein (251 residues) is essential for mini-F replication in Escherichia coli and exhibits two major functions: initiation of DNA replication from ori2 and autogenous repression of repE transcription. Whereas the initiation is mediated by RepE monomers that bind to the ori2 iterons (direct repeats), the autogenous repression is mediated by dimers that bind to the repE operator, which contains an inverted repeat sequence related to the iterons. We now report that the binding of RepE to these DNA sites is primarily determined by the C-terminal region of this protein. The mutant RepE proteins lacking either the N-terminal 33 (or more) residues or the C-terminal 7 (or more) residues were first shown to be defective in binding to both the ori2 and the operator DNAs. However, direct screening and analysis of mutant RepEs which are specifically affected in binding to the ori2 iterons revealed that the mutations (mostly amino acid substitutions) occur exclusively in the C-terminal region (residues 168 to 242). These mutant proteins exhibited reduced binding to ori2 and no detectable binding to the operator. Thus, whereas truncation of either end of RepE can destroy the DNA-binding activities, the C-terminal region appears to represent a primary DNA-binding domain of RepE for both ori2 and the operator. Analogous DNA-binding domains seem to be conserved among the initiator proteins of certain related plasmids.
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含有 iteron 的 DNA 分子复制的调节。
DOI：
10.1016/s0079-6603(08)60857-0
发表时间：
1994
期刊：
Progress in nucleic acid research and molecular biology
影响因子：
--
作者：
Filutowicz,M;Dellis,S;Levchenko,I;Urh,M;Wu,F;York,D
通讯作者：
York,D
质粒 R6K 复制起始蛋白的一级结构。
DOI：
10.1073/pnas.79.18.5475
发表时间：
1982
影响因子：
11.1
作者：
Germino,J;Bastia,D
通讯作者：
Bastia,D
大肠杆菌 F 质粒的自主控制机制：repE 基因产物的纯化和表征。
影响因子：
14.9
作者：
Masson,L;Ray,DS
通讯作者：
Ray,DS
通过基因融合和位点特异性蛋白水解快速纯化克隆基因产物。
DOI：
10.1073/pnas.81.15.4692
发表时间：
1984
影响因子：
11.1
作者：
Germino,J;Bastia,D
通讯作者：
Bastia,D
大肠杆菌 F 质粒的自主控制机制：启动子/阻遏蛋白的不同复合物与其操纵子和 F 质粒复制起点结合。
影响因子：
14.9
作者：
Masson,L;Ray,DS
通讯作者：
Ray,DS