HDX-MS and deletion analysis of the type 4 secretion system protein TraF from the Escherichia coli F plasmid

FEBS Lett. 2016 Feb;590(3):376-86. doi: 10.1002/1873-3468.12066. Epub 2016 Feb 3.

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.

Keywords: F-plasmid; Hydrogen-deuterium exchange; conjugation; thioredoxin fold; type IV secretion.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / metabolism
  • Conjugation, Genetic
  • Deuterium Exchange Measurement
  • Escherichia coli / physiology*
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism*
  • Fimbriae, Bacterial / physiology*
  • Gene Deletion
  • Gene Knockout Techniques
  • Homologous Recombination
  • Kinetics
  • Models, Molecular*
  • Molecular Sequence Data
  • Nuclear Proteins / chemistry
  • Nuclear Proteins / metabolism
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / metabolism
  • Plasmids / chemistry
  • Plasmids / metabolism
  • Protein Conformation
  • Protein Interaction Domains and Motifs
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / metabolism
  • Spectrometry, Mass, Electrospray Ionization
  • Type IV Secretion Systems / chemistry
  • Type IV Secretion Systems / genetics
  • Type IV Secretion Systems / metabolism*

Substances

  • Bacterial Proteins
  • Escherichia coli Proteins
  • Nuclear Proteins
  • Peptide Fragments
  • Recombinant Fusion Proteins
  • TraF protein, E coli
  • TraH protein, Bacteria
  • Type IV Secretion Systems

Associated data

  • GENBANK/BAA97961