Abstract
The Streptomyces virginiae γ-butyrolactone autoregulator virginiae butanolide is a low-molecular-weight Streptomyces hormone eliciting virginiamycin biosynthesis through its binding to the specific receptor protein, BarA. Immediately downstream of barA lies barB, the transcription of which is tightly repressed by BarA in the absence of virginiae butanolide and derepressed in its presence. Thus, BarB is next to BarA on the virginiae butanolide-BarA signaling cascade. An in-frame 279-bp deletion was introduced into the barB allele, which rendered it inactive by eliminating the majority of the coding region, including the helix-turn-helix DNA-binding motif. No significant change was observed with the ΔbarB mutant with respect to the timing or amount of virginiae butanolide production, or the morphological differentiation on solid media, indicating that barB neither participates in virginiae butanolide biosynthesis nor in cytodifferentiation. In contrast, analysis of virginiamycin production in the ΔbarB mutant revealed that production of both virginiamycin M1 and virginiamycin S occurred immediately after virginiae butanolide production, 2–3 h earlier than in the wild-type strain, indicating that BarB participates in the temporal retardation of virginiamycin production after virginiae butanolide inactivates the repressor function of BarA. RT-PCR analysis of the transcription of several genes surrounding barA–barB by the ΔbarB mutant indicated that BarB plays a negative regulatory role, directly or indirectly, in the transcription of barZ, vmsR, and orf5 located upstream of barB.
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Acknowledgements
This work was supported in part by grants from the Research for the Future Program of the Japan Society for the Promotion of Science (JSPS), the National Food Research Institute (Tsukuba, Japan), and the Korean Research Foundation (KRF-2002-C00086). KM was a fellow of JSPS.
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Matsuno, K., Yamada, Y., Lee, CK. et al. Identification by gene deletion analysis of barB as a negative regulator controlling an early process of virginiamycin biosynthesis in Streptomyces virginiae . Arch Microbiol 181, 52–59 (2004). https://doi.org/10.1007/s00203-003-0625-5
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DOI: https://doi.org/10.1007/s00203-003-0625-5