title: HDX-MS and deletion analysis of the type 4 secretion system protein TraF from the Escherichia coli F plasmid.
abstract: Conjugative DNA transfer by the F-plasmid is achieved through a type IV secretion system (T4SS) encoded within the plasmid's transfer region; TraF is one of several F-T4SS proteins essential for F-pilus assembly. In order to identify regions of the protein important for TraF function, a series of deletion mutants were assessed for their ability to recover conjugative transfer in a traF knockout. Interestingly, modification of any region of TraF abolishes pilus synthesis, resulting in a loss of rescue of conjugative function. Dynamic analysis of TraF by time-resolved hydrogen-deuterium exchange revealed that the C-terminal region containing the predicted thioredoxin-like domain is quite structured, while the N-terminal region, predicted to interact with TraH in the intact F-T4SS, was more dynamic.
authors.affiliation: Department of Chemistry, York University, Toronto, ON, Canada.
authors.forename: Cristina
authors.initials: C
authors.lastname: Lento
authors.affiliation: Department of Chemistry, York University, Toronto, ON, Canada.
authors.forename: Michele
authors.initials: M
authors.lastname: Ferraro
authors.affiliation: Department of Chemistry, York University, Toronto, ON, Canada.|Centre for Research on Biomolecular Interactions, York University, Toronto, ON, Canada.|Centre for Research in Mass Spectrometry, York University, Toronto, ON, Canada.
authors.forename: Derek
authors.identifier: http://orcid.org/0000-0002-7012-6085
authors.initials: D
authors.lastname: Wilson
authors.affiliation: Department of Chemistry, York University, Toronto, ON, Canada.|Centre for Research on Biomolecular Interactions, York University, Toronto, ON, Canada.
authors.forename: Gerald F
authors.identifier: http://orcid.org/0000-0002-5695-9830
authors.initials: GF
authors.lastname: Audette
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D003227:Conjugation, Genetic; D041961:Deuterium Exchange Measurement; D004926:Escherichia coli / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010861:Fimbriae, Bacterial / Q000502:physiology*; D017353:Gene Deletion; D055786:Gene Knockout Techniques; D059765:Homologous Recombination; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D000069319:Type IV Secretion Systems / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D009687:Nuclear Proteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C500309:TraF protein, E coli; C065598:TraH protein, Bacteria; D000069319:Type IV Secretion Systems
keywords: F-plasmid; Hydrogen-deuterium exchange; conjugation; thioredoxin fold; type IV secretion