The putative coupling protein TcpA interacts with other pCW3-encoded proteins to form an essential part of the conjugation complex

Jennifer Anne Steen, Trudi Leanne Bannam, Wee Lin Teng, Rodney James Devenish, Julian Ian Rood

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Conjugative plasmids encode antibiotic resistance determinants or toxin genes in the anaerobic pathogen Clostridium perfringens. The paradigm conjugative plasmid in this bacterium is pCW3, a 47kb tetracycline resistance plasmid that encodes the unique tcp transfer locus. The tcp locus consists of 11 genes, intP and tcpA-tcpJ, at least three of which, tcpA, tcpF and tcpH, 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.
Original languageEnglish
Pages (from-to)2926 - 2933
Number of pages8
JournalJournal of Bacteriology
Issue number9
Publication statusPublished - 2009

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