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.
© 2016 Federation of European Biochemical Societies.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Bacterial Proteins / chemistry
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Bacterial Proteins / metabolism
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Conjugation, Genetic
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Deuterium Exchange Measurement
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Escherichia coli / physiology*
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Escherichia coli Proteins / chemistry
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Escherichia coli Proteins / genetics
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Escherichia coli Proteins / metabolism*
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Fimbriae, Bacterial / physiology*
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Gene Deletion
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Gene Knockout Techniques
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Homologous Recombination
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Kinetics
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Models, Molecular*
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Molecular Sequence Data
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Nuclear Proteins / chemistry
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Nuclear Proteins / metabolism
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Peptide Fragments / chemistry
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Peptide Fragments / genetics
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Peptide Fragments / metabolism
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Plasmids / chemistry
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Plasmids / metabolism
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Protein Conformation
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Protein Interaction Domains and Motifs
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / metabolism
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Spectrometry, Mass, Electrospray Ionization
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Type IV Secretion Systems / chemistry
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Type IV Secretion Systems / genetics
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Type IV Secretion Systems / metabolism*
Substances
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Bacterial Proteins
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Escherichia coli Proteins
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Nuclear Proteins
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Peptide Fragments
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Recombinant Fusion Proteins
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TraF protein, E coli
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TraH protein, Bacteria
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Type IV Secretion Systems