Abstract
Myo-inositol is important for Streptomyces growth and morphological differentiation. Genomic sequence analysis revealed a myo-inositol catabolic gene cluster in Streptomyces coelicolor. Disruption of the corresponding genes in this cluster abolished the bacterial growth on myo-inositol as a single carbon source. The transcriptions of these genes were remarkably enhanced by addition of myo-inositol in minimal medium. A putative regulatory gene SCO6974, encoding a GntR family protein, is situated in the cluster. Disruption of SCO6974 significantly enhanced the transcription of myo-inositol catabolic genes. SCO6974 was shown to interact with the promoter regions of myo-inositol catabolic genes using electrophoretic mobility shift assays. DNase I footprinting assays demonstrated that SCO6974 recognized a conserved palindromic sequence (A/T)TGT(A/C)N(G/T)(G/T)ACA(A/T). Base substitution of the conserved sequence completely abolished the binding of SCO6974 to the targets demonstrating that SCO6974 directly represses the transcriptions of myo-inositol catabolic genes. Furthermore, the disruption of SCO6974 was correlated with a reduced sporulation of S. coelicolor in mannitol soya flour medium and with the overproduction of actinorhodin and calcium-dependent antibiotic. The addition of myo-inositol suppressed the sporulation deficiency of the mutant, indicating that the effect could be related to a shortage in myo-inositol due to its enhanced catabolism in this strain. This enhanced myo-inositol catabolism likely yields dihydroxyacetone phosphate and acetyl-CoA that are indirect or direct precursors of the overproduced antibiotics.
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Acknowledgments
This work was supported by grants from the National Natural Science Foundation of China (grant numbers 31170088 and 31200929). We are grateful to Prof. Keith Chater (John Innes Centre, Norwich, UK) for providing E. coli ET12567/pUZ8002 and plasmid pSET152. We would like to thank Dr. Ying Wen (China Agriculture University, Beijing, China) for her kind help in EMSAs.
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L.Y. and S.L. contributed equally to this work.
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Yu, L., Li, S., Gao, W. et al. Regulation of myo-inositol catabolism by a GntR-type repressor SCO6974 in Streptomyces coelicolor . Appl Microbiol Biotechnol 99, 3141–3153 (2015). https://doi.org/10.1007/s00253-014-6368-1
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DOI: https://doi.org/10.1007/s00253-014-6368-1