The Vibrio cholerae fatty acid regulatory protein, FadR, represses transcription of plsB, the gene encoding the first enzyme of membrane phospholipid biosynthesis

Youjun Feng, John E. Cronan

Research output: Contribution to journalArticlepeer-review

Abstract

Glycerol-3-phosphate (sn-glycerol-3-P, G3P) acyltransferase catalyses the first committed step in the biosynthesis of membrane phospholipids, the acylation of G3P to form 1-acyl G3P (lysophosphatidic acid). The paradigm G3P acyltransferase is the Escherichia coli plsB gene product which acylates position-1 of G3P using fatty acids in thioester linkage to either acyl carrier protein (ACP) or CoA as acyl donors. Although the E.coli plsB gene was discovered about 30 years ago, no evidence for transcriptional control of its expression has been reported. However A.E. Kazakov and co-workers (J Bacteriol 2009; 191: 52-64) reported the presence of a putative FadR binding site upstream of the candidate plsB genes of Vibrio cholerae and three other Vibrio species suggesting that plsB might be regulated by FadR, a GntR family transcription factor thus far known only to regulate fatty acid synthesis and degradation. We report that the V.cholerae plsB homologue restored growth of E.coli strain BB26-36 which is a G3P auxotroph due to an altered G3P acyltransferase activity. The plsB promoter was also mapped and the predicted FadR-binding palindrome was found to span positions -19 to -35, upstream of the transcription start site. Gel shift assays confirmed that both V.cholerae FadR and E.coli FadR bound the V.cholerae plsB promoter region and binding was reversed upon addition of long-chain fatty acyl-CoA thioesters. The expression level of the V.cholerae plsB gene was elevated two- to threefold in an E.coli fadR null mutant strain indicating that FadR acts as a repressor of V.cholerae plsB expression. In both E.coli and V.cholerae the β-galactosidase activity of transcriptional fusions of the V.cholerae plsB promoter to lacZ increased two- to threefold upon supplementation of growth media with oleic acid. Therefore, V.cholerae co-ordinates fatty acid metabolism with 1-acyl G3P synthesis.

Original languageEnglish (US)
Pages (from-to)1020-1033
Number of pages14
JournalMolecular Microbiology
Volume81
Issue number4
DOIs
StatePublished - Aug 2011

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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