Characterization of the srfA locus of Bacillus subtilis: only the valine-activating domain of srfA is involved in the establishment of genetic competence.
srfA is a locus required for the production of the lipopeptide antibiotic surfactin. This locus is also necessary for efficient sporulation and competence development. Mutations in the 5' portion of the srfA operon affect all three of these processes, whereas mutations in the 3' portion of srfA only affect sporulation and surfactin production. Analysis of the proteins encoded by the srfA locus revealed seven large domains which are likely to be responsible for the activation and binding of the seven amino acids of surfactin. Identification of the amino acid that is activated by the srfA domains was determined by amino acid-dependent pyrophosphate exchange reactions on partially purified cell extracts of strains carrying different srfA mutations. These results indicate colinearity between the order of the domains in the srfA locus and the amino acid sequence of surfactin. The minimal genetic element of srfA required for the establishment of competence was shown to be the 5' region of the second open reading of srfA, which encodes the valine activation domain. This portion of srfA, when cloned on a plasmid, complemented the competence deficiency of a srfA deletion mutant in trans.
van Sinderen
D
D
Department of Genetics, Centre of Biological Sciences, Haren Groningen, The Netherlands.
Galli
G
G
Cosmina
P
P
de Ferra
F
F
Withoff
S
S
Venema
G
G
Grandi
G
G
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics* / Q000502:physiology; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D001665:Binding Sites; D015964:Gene Expression Regulation, Bacterial; D055666:Lipopeptides; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010453:Peptide Synthases / Q000737:chemistry* / Q000235:genetics; D010456:Peptides, Cyclic; D017434:Protein Structure, Tertiary; D015398:Signal Transduction / Q000235:genetics*; D013171:Spores, Bacterial; D014169:Transformation, Bacterial / Q000235:genetics*; D014633:Valine
D001426:Bacterial Proteins; D055666:Lipopeptides; D010456:Peptides, Cyclic; C009365:surfactin peptide; D010453:Peptide Synthases; C069542:surfactin synthetase; D014633:Valine