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Control of growth, secondary metabolism and sporulation in Streptomyces venezuelae ISP5230 by jadW₁, a member of the afsA family of -butyrolactone regulatory genes

Department of Biology, Dalhousie University, Halifax, Nova Scotia, Canada B3H 4J1

Correspondence
Leo C. Vining
leo.vining{at}dal.ca

Three new genes (jadW₁, jadW₂ and jadW₃) were isolated from a region of the Streptomyces venezuelae ISP5230 chromosome at the left-hand end of the jad cluster for jadomycin B (JdB) biosynthesis. The deduced amino acid sequence of jadW₁ showed strong similarity to gene products associated in several streptomycetes with -butyrolactone autoregulators controlling morphological differentiation and secondary metabolism. Examination of JadW₁ for conserved domains detected a repeat sequence characteristic of proteins in the AfsA regulatory family. Insertional inactivation of jadW₁ reduced the growth rate of S. venezuelae cultures in aerated liquid media containing complex nitrogen sources and altered growth morphology in minimal medium. It also affected sporulation on agar media. Cultures of jadW₁-disrupted mutants grown under conditions supporting biosynthesis of JdB or chloramphenicol by the wild-type strain failed to produce either of the antibiotics. Complementing the disrupted strain by transformation with pJV435, containing a cloned copy of the gene, improved sporulation and restored antibiotic biosynthesis in transformants to titres close to those of the wild-type similarly transformed with pJV435 as a control. The results are consistent with a role for jadW₁ in regulating morphological and metabolic differentiation. Further sequence analysis of jadR₂, which functions with jadR₁ in stress-induced activation of JdB biosynthesis, indicated that this gene encodes a -butyrolactone receptor homologue. The growth-rate-sensitive phenotype of the jadW₁-disrupted mutant, and the proximity of jadW₁ to jadR₂ indicate that this region of the jad gene cluster contains a regulatory mechanism incorporating -butyrolactone signalling and sensitivity to environmental stress.

The GenBank accession number for the sequence reported in this paper is U24659.

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