期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 105, 期 36, 页码 13321-13326出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0806873105
关键词
antibiotic resistance; fatty acid biosynthesis inhibitor
资金
- National Institute of Health [GM20011]
- Swiss National Science Foundation
- Ernst Schering Foundation
Andrimid is a hybrid nonribosomal peptide-polyketide antibiotic that blocks the carboxyl-transfer reaction of bacterial acetyl-CoA carboxylase (ACC) and thereby inhibits fatty acid biosynthesis with submicromolar potency. The andrimid biosynthetic gene cluster from Pantoea agglomerans encodes an admT gene with homology to the acetyl-CoA carboxyltransferase (CT) beta-subunit gene accD. Escherichia coli cells overexpressing admT showed resistance to andrimid. Co-overproduction of AdmT with E. coli CT alpha-subunit AccA allowed for the in vitro reconstitution of an active heterologous tetrameric CT A(2)T(2) complex. A subsequent andrimid-inhibition assay revealed an IC50 of 500 nM for this hybrid A2T2 in contrast to that of 12 nM for E. coli CT A(2)D(2). These results validated that AdmT is an AccD homolog that confers resistance in the andrimid producer. Mutagenesis studies guided by the x-ray crystal structure of the E. coli A2D2 complex disclosed a single amino acid mutation of AdmT (L203M) responsible for 5-fold andrimid sensitivity (IC50 = 100 nM). Complementarily, the E. coli AccD mutant M203L became 5-fold more resistant in the CT assays. This observation allowed for bioinformatic identification of several Vibrio cholerae strains in which accD genes encode the Met <-> Leu switches, and their occurrences correlate predictively with sensitivities to andrimid in vivo.
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