title: The mating pair stabilization protein, TraN, of the F plasmid is an outer-membrane protein with two regions that are important for its function in conjugation.
abstract: F plasmid TraN (602 aa, processed to 584 aa with 22 conserved cysteines), which is essential for F plasmid conjugation, is an outer-membrane protein involved in mating pair stabilization (MPS). Unlike R100 TraN, F TraN requires OmpA in the recipient cell for efficient MPS. The authors have identified three external loops (aa 172-187, 212-220 and 281-284) in the highly divergent region from aa 164 to aa 333 as candidates for interaction with OmpA. These loops were identified using both site-directed and random TnphoA/in mutagenesis to insert epitopes (31-aa or c-myc) into TraN and monitor their effect on sensitivity to external proteases and on mating ability. TraN is a hallmark protein of F-type IV secretion systems as demonstrated by blast searches of the databases. The C-terminal region is highly conserved and contains five of the six completely conserved cysteines. Mutation of these residues to serine demonstrated their importance in TraN function. TraN appears to require both intra- and intermolecular disulfide bond formation for its stability and structure as demonstrated by its instability in a dsbA mutant and its aberrant migration on SDS-polyacrylamide gels under non-reducing conditions or by cross-linking with bis(sulfosuccinimidyl)suberate (BS3). Thus, F TraN appears to have two domains: the N-terminal region is involved in OmpA interaction with OmpA during MPS; and the C-terminal region, which is rich in conserved cysteine residues, is essential for conjugation.
authors.affiliation: CW405, Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada T6G 2E9.
authors.forename: William A
authors.initials: WA
authors.lastname: Klimke
authors.affiliation: CW405, Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada T6G 2E9.
authors.forename: Candace D
authors.initials: CD
authors.lastname: Rypien
authors.affiliation: CW405, Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada T6G 2E9.
authors.forename: Barbara
authors.initials: B
authors.lastname: Klinger
authors.affiliation: CW405, Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada T6G 2E9.
authors.forename: R Alexander
authors.initials: RA
authors.lastname: Kennedy
authors.affiliation: CW405, Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada T6G 2E9.
authors.forename: J Manuel
authors.initials: JM
authors.lastname: Rodriguez-Maillard
authors.affiliation: CW405, Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada T6G 2E9.
authors.forename: Laura S
authors.initials: LS
authors.lastname: Frost
mesh_terms: D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003227:Conjugation, Genetic; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D005144:F Factor / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional
chemical_list: D001425:Bacterial Outer Membrane Proteins; D004251:DNA Transposable Elements; D029968:Escherichia coli Proteins; C074745:traN protein, E coli