The putative coupling protein TcpA interacts with other pCW3-encoded proteins to form an essential part of the conjugation complex.
The putative coupling protein TcpA interacts with other pCW3-encoded proteins to form an essential part of the conjugation complex.
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假定的偶联蛋白 TcpA 与其他 pCW3 编码蛋白相互作用，形成缀合复合物的重要组成部分。
DOI：
10.1128/jb.00032-09
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发表时间：
2009
影响因子：
3.2
通讯作者：
Rood,JulianI
中科院分区：
文献类型：
--
作者：
Steen,JenniferA;Bannam,TrudiL;Teng,WeeLin;Devenish,RodneyJ;Rood,JulianI
接合质粒编码厌氧病原体产气荚膜梭菌中的抗生素抗性决定簇或毒素基因。该细菌中的典型接合质粒是pCW 3，其是编码四环素转移位点的47-kb四环素抗性质粒。tcplocus由intPandtcpA-tcpJ共11个基因组成，其中至少有三个基因tcpA、tcpF和tcpH是pCW 3接合转移所必需的。在这项研究中，我们研究了蛋白质-蛋白质相互作用涉及TcpA，推定的耦合蛋白。使用细菌双杂交系统确定TcpA和TcpC，TcpG和TcpH之间的相互作用。该分析还证明了TcpA、TcpC和TcpG自身相互作用，这通过化学交联研究得到证实。对TcpA的一系列缺失和定点衍生物的检查确定了这些相互作用所需的结构域和基序。基于这些结果，我们已经构建了一个模型，这种独特的共轭转移装置。
Conjugative plasmids encode antibiotic resistance determinants or toxin genes in the anaerobic pathogenClostridium perfringens. The paradigm conjugative plasmid in this bacterium is pCW3, a 47-kb tetracycline resistance plasmid that encodes the uniquetcptransfer locus. Thetcplocus consists of 11 genes,intPandtcpA-tcpJ, at least three of which,tcpA,tcpF, andtcpH, are essential for the conjugative transfer of pCW3. In this study we examined protein-protein interactions involving TcpA, the putative coupling protein. Use of a bacterial two-hybrid system identified interactions between TcpA and TcpC, TcpG, and TcpH. This analysis also demonstrated TcpA, TcpC, and TcpG self-interactions, which were confirmed by chemical cross-linking studies. Examination of a series of deletion and site-directed derivatives of TcpA identified the domains and motifs required for these interactions. Based on these results, we have constructed a model for this unique conjugative transfer apparatus.