A 20-residue peptide of the inner membrane protein OutC mediates interaction with two distinct sites of the outer membrane secretin OutD and is essential for the functional type II secretion system in Erwinia chrysanthemi.
The type II secretion system (T2SS) is widely exploited by proteobacteria to secrete enzymes and toxins involved in bacterial survival and pathogenesis. The outer membrane pore formed by the secretin OutD and the inner membrane protein OutC are two key components of the secretion complex, involved in secretion specificity. Here, we show that the periplasmic regions of OutC and OutD interact directly and map the interaction site of OutC to a 20-residue peptide named OutCsip (secretin interacting peptide, residues 139-158). This peptide interacts in vitro with two distinct sites of the periplasmic region of OutD, one located on the N0 subdomain and another overlapping the N2-N3' subdomains. The two interaction sites of OutD have different modes of binding to OutCsip. A single substitution, V143S, located within OutCsip prevents its interaction with one of the two binding sites of OutD and fully inactivates the T2SS. We show that the N0 subdomain of OutD interacts also with a second binding site within OutC located in the region proximal to the transmembrane segment. We suggest that successive interactions between these distinct regions of OutC and OutD may have functional importance in switching the secretion machine.
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Frédéric H
FH
Université de Lyon, F-69003, Université Lyon 1, Lyon, F-69622, INSA-Lyon, Villeurbanne, F-69621, CNRS, UMR5240, Microbiologie Adaptation et Pathogénie, Lyon, F-69622, France.
Fries
Markus
M
Wang
Xiaohui
X
Pickersgill
Richard W
RW
Shevchik
Vladimir E
VE
D019943:Amino Acid Substitution; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D016974:Dickeya chrysanthemi / Q000378:metabolism*; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D025941:Protein Interaction Mapping; D021381:Protein Transport
D001425:Bacterial Outer Membrane Proteins; C105759:OutC protein, bacteria; C106887:OutD protein, Erwinia chrysanthemi; D010455:Peptides