PMID:22150951

Citation	Porter, CJ, Bantwal, R, Bannam, TL, Rosado, CJ, Pearce, MC, Adams, V, Lyras, D, Whisstock, JC and Rood, JI (2012) The conjugation protein TcpC from Clostridium perfringens is structurally related to the type IV secretion system protein VirB8 from Gram-negative bacteria. Mol. Microbiol. 83:275-88
Abstract	Bacterial conjugation is important for the acquisition of virulence and antibiotic resistance genes. We investigated the mechanism of conjugation in Gram-positive pathogens using a model plasmid pCW3 from Clostridium perfringens. pCW3 encodes tetracycline resistance and contains the tcp locus, which is essential for conjugation. We showed that the unique TcpC protein (359 amino acids, 41 kDa) was required for efficient conjugative transfer, localized to the cell membrane independently of other conjugation proteins, and that membrane localization was important for its function, oligomerization and interaction with the conjugation proteins TcpA, TcpH and TcpG. The crystal structure of the C-terminal component of TcpC (TcpC(99-359)) was determined to 1.8-Å resolution. TcpC(99-359) contained two NTF2-like domains separated by a short linker. Unexpectedly, comparative structural analysis showed that each of these domains was structurally homologous to the periplasmic region of VirB8, a component of the type IV secretion system from Agrobacterium tumefaciens. Bacterial two-hybrid studies revealed that the C-terminal domain was critical for interactions with other conjugation proteins. The N-terminal region of TcpC was required for efficient conjugation, oligomerization and protein-protein interactions. We conclude that by forming oligomeric complexes, TcpC contributes to the stability and integrity of the conjugation apparatus, facilitating efficient pCW3 transfer.
Links	PubMed Online version:10.1111/j.1365-2958.2011.07930.x
Keywords	Agrobacterium tumefaciens/chemistry; Agrobacterium tumefaciens/genetics; Bacterial Proteins/chemistry; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Cell Membrane/chemistry; Clostridium perfringens/chemistry; Clostridium perfringens/genetics; Clostridium perfringens/metabolism; Conjugation, Genetic; Crystallography, X-Ray; Molecular Weight; Plasmids/metabolism; Protein Binding; Protein Interaction Mapping; Protein Multimerization; Protein Structure, Tertiary; Two-Hybrid System Techniques; Virulence Factors/chemistry; Virulence Factors/genetics
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
CLOPF:Q1PLH8		GO:0016020: membrane	ECO:0000314:		C		TcpC was detected in the membrane fraction of the wild-type strain and a complemented tcpC mutant, but was absent in the tcpC mutant carrying the shuttle vector control (Fig. 3). Clostridium perfringens	complete CACAO 12585

References

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