Absence of the outer membrane phospholipase a suppresses the temperature-sensitive phenotype of Escherichia coli degP mutants and induces the Cpx and σEextracytoplasmic stress responses

G. R. Langen, J. R. Harper, T. J. Silhavy, S. P. Howard

Research output: Contribution to journalArticle

19 Scopus citations

Abstract

DegP is a periplasmic protease that is a member of both the σEand Cpx extracytoplasmic stress regulons of Escherichia coli and is essential for viability at temperatures above 42°C. [U-14C]acetate labeling experiments demonstrated that phospholipids were degraded in degP mutants at elevated temperatures. In addition, chloramphenicol acetyltransferase, β-lactamase, and β-galactosidase assays as well as sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis indicated that large amounts of cellular proteins are released from degP cells at the nonpermissive temperature. A mutation in pldA, which encodes outer membrane phospholipase A (OMPLA), was found to rescue degP cells from the temperature-sensitive phenotype, pldA degP mutants had a normal plating efficiency at 42°C, displayed increased viability at 44°C, showed no degradation of phospholipids, and released far lower amounts of cellular protein to culture supernatants. degP and pldA degP mutants containing chromosomal lacZ fusions to Cpx and σEregulon promoters indicated that both regulons were activated in the pldA mutants. The overexpression of the envelope lipoprotein, NlpE, which induces the Cpx regulon, was also found to suppress the temperature-sensitive phenotype of degP mutants but did not prevent the degradation of phospholipids. These results suggest that the absence of OMPLA corrects the degP temperature-sensitive phenotype by inducing the Cpx and σE regulons rather than by inactivating the phospholipase per se.

Original languageEnglish (US)
Pages (from-to)5230-5238
Number of pages9
JournalJournal of bacteriology
Volume183
Issue number18
DOIs
StatePublished - 2001
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology

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