A nonribosomal peptide synthetase with a novel domain organization is essential for siderophore biosynthesis in Vibrio anguillarum.
Anguibactin, a siderophore produced by Vibrio anguillarum, is synthesized via a nonribosomal peptide synthetase (NRPS) mechanism. We have identified a gene from the V. anguillarum plasmid pJM1 that encodes a 78-kDa NRPS protein termed AngM, which is essential in the biosynthesis of anguibactin. The predicted AngM amino acid sequence shows regions of homology to the consensus sequence for the peptidyl carrier protein (PCP) and the condensation (C) domains of NRPSs, and curiously, these two domains are not associated with an adenylation (A) domain. Substitution by alanine of the serine 215 in the PCP domain and of histidine 406 in the C domain of AngM results in an anguibactin-deficient phenotype, underscoring the importance of these two domains in the function of this protein. The mutations in angM that affected anguibactin production also resulted in a dramatic attenuation of the virulence of V. anguillarum 775, highlighting the importance of this gene in the establishment of a septicemic infection in the vertebrate host. Transcription of the angM gene is initiated at an upstream transposase gene promoter that is repressed by the Fur protein in the presence of iron. Analysis of the sequence at this promoter showed that it overlaps the iron transport-biosynthesis promoter and operates in the opposite direction.
Di Lorenzo
Manuela
M
Department of Molecular Microbiology and Immunology, Oregon Health and Science University, Portland, OR 97201, USA.
Poppelaars
Sophie
S
Stork
Michiel
M
Nagasawa
Maho
M
Tolmasky
Marcelo E
ME
Crosa
Jorge H
JH
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D004251:DNA Transposable Elements; D005393:Fish Diseases / Q000382:microbiology; D015964:Gene Expression Regulation, Bacterial; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D017686:Oncorhynchus mykiss / Q000382:microbiology*; D010453:Peptide Synthases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010455:Peptides / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D014733:Vibrio / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity*; D014735:Vibrio Infections / Q000382:microbiology / Q000662:veterinary
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D010455:Peptides; C049469:anguibactin; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase