title: The conjugation protein TcpC from Clostridium perfringens is structurally related to the type IV secretion system protein VirB8 from Gram-negative bacteria.
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.
authors.affiliation: ARC Centre of Excellence in Structural and Functional Microbial Genomics, Clayton, Vic. 3800, Australia.
authors.forename: Corrine J
authors.initials: CJ
authors.lastname: Porter
authors.forename: Radhika
authors.initials: R
authors.lastname: Bantwal
authors.forename: Trudi L
authors.initials: TL
authors.lastname: Bannam
authors.forename: Carlos J
authors.initials: CJ
authors.lastname: Rosado
authors.forename: Mary C
authors.initials: MC
authors.lastname: Pearce
authors.forename: Vicki
authors.initials: V
authors.lastname: Adams
authors.forename: Dena
authors.initials: D
authors.lastname: Lyras
authors.forename: James C
authors.initials: JC
authors.lastname: Whisstock
authors.forename: Julian I
authors.initials: JI
authors.lastname: Rood
mesh_terms: D016960:Agrobacterium tumefaciens / Q000737:chemistry / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry; D003016:Clostridium perfringens / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D003227:Conjugation, Genetic; D018360:Crystallography, X-Ray; D008970:Molecular Weight; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D025941:Protein Interaction Mapping; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D020798:Two-Hybrid System Techniques; D037521:Virulence Factors / Q000737:chemistry / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D037521:Virulence Factors