Identification by gene deletion analysis of barB as a negative regulator controlling an early process of virginiamycin biosynthesis in Streptomyces virginiae.
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 Delta barB 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 Delta barB 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 Delta barB mutant indicated that BarB plays a negative regulatory role, directly or indirectly, in the transcription of barZ, vmsR, and orf5 located upstream of barB.
Matsuno
Kiyoshi
K
International Center for Biotechnology, Osaka University, 2--1 Yamadaoka, Suita, 565-0871 Osaka, Japan.
Yamada
Yasuhiro
Y
Lee
Chang-Kwon
CK
Nihira
Takuya
T
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology*; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D023061:Gene Order; D005798:Genes, Bacterial; D019077:Helix-Turn-Helix Motifs / Q000235:genetics; D008969:Molecular Sequence Data; D020161:Physical Chromosome Mapping; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D012097:Repressor Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D025364:Streptogramin A / Q000096:biosynthesis; D025382:Streptogramin Group B / Q000096:biosynthesis; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic / Q000502:physiology; D014769:Virginiamycin / Q000096:biosynthesis*
D001426:Bacterial Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins; D025382:Streptogramin Group B; D014769:Virginiamycin; D025364:Streptogramin A; C418083:virginiamycin factor S1