Journal
ARCHIVES OF MICROBIOLOGY
Volume 181, Issue 1, Pages 52-59Publisher
SPRINGER
DOI: 10.1007/s00203-003-0625-5
Keywords
Streptomyces virginiae; gamma-butyrolactone autoregulator; virginiamycin production; repressor BarB; gene disruption
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The Streptomyces virginiae gamma-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 DeltabarB 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 DeltabarB mutant revealed that production of both virginiamycin M-1 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 DeltabarB 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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