title: Identification of YidC residues that define interactions with the Sec Apparatus.
abstract: In bacteria, a subset of membrane proteins insert into the membrane via the Sec apparatus with the assistance of the widely conserved essential membrane protein insertase YidC. After threading into the SecYEG translocon, transmembrane segments of nascent proteins are thought to exit the translocon via a lateral gate in SecY, where YidC facilitates their transfer into the lipid bilayer. Interactions between YidC and components of the Sec apparatus are critical to its function. The first periplasmic loop of YidC interacts directly with SecF. We sought to identify the regions or residues of YidC that interact with SecY or with additional components of the Sec apparatus other than SecDF. Using a synthetic lethal screen, we identified residues of YidC that, when mutated, led to dependence on SecDF for viability. Each residue identified is highly conserved among YidC homologs; most lie within transmembrane domains. Overexpression of SecY in the presence of two YidC mutants partially rescued viability in the absence of SecDF, suggesting that the corresponding wild-type YidC residues (G355 and M471) participate in interactions, direct or indirect, with SecY. Staphylococcus aureus YidC complemented depletion of YidC, but not of SecDF, in Escherichia coli. G355 of E. coli YidC is invariant in S. aureus YidC, suggesting that this highly conserved glycine serves a conserved function in interactions with SecY. This study demonstrates that transmembrane residues are critical in YidC interactions with the Sec apparatus and provides guidance on YidC residues of interest for future structure-function analyses.
authors.affiliation: Division of Infectious Diseases, Department of Medicine, Massachusetts General Hospital, Cambridge, Massachusetts, USA.
authors.forename: Zaoping
authors.initials: Z
authors.lastname: Li
authors.forename: Dana
authors.initials: D
authors.lastname: Boyd
authors.forename: Martin
authors.initials: M
authors.lastname: Reindl
authors.forename: Marcia B
authors.initials: MB
authors.lastname: Goldberg
mesh_terms: D004252:DNA Mutational Analysis; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism*; D005816:Genetic Complementation Test; D026901:Membrane Transport Proteins / Q000235:genetics* / Q000378:metabolism*; D050296:Microbial Viability; D025941:Protein Interaction Mapping; D013211:Staphylococcus aureus / Q000235:genetics
chemical_list: D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; C412443:YIDC protein, E coli