Assembly-defective OmpC mutants of Escherichia coli K-12.
Novel ompC(Dex) alleles were utilized to isolate mutants defective in OmpC biogenesis. These ompC(Dex) alleles also conferred sensitivity to sodium dodecyl sulfate (SDS), which permitted the isolation of SDS-resistant and OmpC-specific phage-resistant mutants that remained Dex+. Many mutants acquired resistance against these lethal agents by lowering the OmpC level present in the outer membrane. In the majority of these mutants, a defect in the assembly (metastable to stable trimer formation) was responsible for lowering OmpC levels. The assembly defects in various mutant OmpC proteins were caused by single-amino-acid substitutions involving the G-39, G-42, G-223, G-224, Q-240, G-251, and G-282 residues of the mature protein. This assembly defect was correctable by an assembly suppressor allele, asmA3. In addition, we investigated one novel OmpC mutant in which an assembly defect was caused by a disulfide bond formation between two nonnative cysteine residues. The assembly defect was fully corrected in a genetic background in which the cell's ability to form disulfide bonds was compromised. The assembly defect of the two-cysteine OmpC protein was also mended by asmA3, whose suppressive effect was not achieved by preventing disulfide bond formation in the mutant OmpC protein.
Xiong
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Department of Microbiology, Arizona State University, Tempe 85287-2701, USA.
Deeter
J N
JN
Misra
R
R
D000483:Alleles; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics; D003090:Coliphages / Q000472:pathogenicity; D003545:Cysteine / Q000235:genetics; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000821:virology; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D009154:Mutation; D010641:Phenotype; D011134:Polysaccharides / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D013489:Suppression, Genetic
C096310:AsmA protein, E coli; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D011134:Polysaccharides; D012967:Sodium Dodecyl Sulfate; C008315:maltodextrin; D003545:Cysteine