PMID:27103580

Citation	Fercher, C, Probst, I, Kohler, V, Goessweiner-Mohr, N, Arends, K, Grohmann, E, Zangger, K, Meyer, NH and Keller, W (2016) VirB8-like protein TraH is crucial for DNA transfer in Enterococcus faecalis. Sci Rep 6:24643
Abstract	Untreatable bacterial infections caused by a perpetual increase of antibiotic resistant strains represent a serious threat to human healthcare in the 21(st) century. Conjugative DNA transfer is the most important mechanism for antibiotic resistance and virulence gene dissemination among bacteria and is mediated by a protein complex, known as type IV secretion system (T4SS). The core of the T4SS is a multiprotein complex that spans the bacterial envelope as a channel for macromolecular secretion. We report the NMR structure and functional characterization of the transfer protein TraH encoded by the conjugative Gram-positive broad-host range plasmid pIP501. The structure exhibits a striking similarity to VirB8 proteins of Gram-negative secretion systems where they play an essential role in the scaffold of the secretion machinery. Considering TraM as the first VirB8-like protein discovered in pIP501, TraH represents the second protein affiliated with this family in the respective transfer operon. A markerless traH deletion in pIP501 resulted in a total loss of transfer in Enterococcus faecalis as compared with the pIP501 wild type (wt) plasmid, demonstrating that TraH is essential for pIP501 mediated conjugation. Moreover, oligomerization state and topology of TraH in the native membrane were determined providing insights in molecular organization of a Gram-positive T4SS.
Links	PubMed PMC4840375 Online version:10.1038/srep24643
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
ENTFL:A0A140UHJ9		GO:0000746: conjugation	ECO:0000315:		P		Supplementary Table 3 and section "TraH is an essential component in pIP501-mediated conjugation." Although the paper didn't use the Enterococcus faecalis strain EnGen0234, the Protein Data Bank accession code mentioned in the paper matches the UniProt ID for the EnGen strain. The GO term is appropriate because a mutant strain with a deletion of the traH gene did not conjugate at a level that could be detected by the assay used. However, when the same strain received a plasmid containing a functional copy of the gene, conjugation proceeded at a normal rate.	complete CACAO 12432

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