Cho Y H, Lee E J, Ahn B E, Roe J H
Laboratory of Molecular Microbiology, School of Biological Sciences, and Institute of Microbiology, Seoul National University, Seoul 151-742, Korea.
Mol Microbiol. 2001 Oct;42(1):205-14. doi: 10.1046/j.1365-2958.2001.02622.x.
A gene (sigB) encoding an alternative sigma factor sigmaB in Streptomyces coelicolor A3(2) was isolated and characterized. It encodes a polypeptide of 281 amino acids (31 546 Da) and is highly homologous to Bacillus subtilis sigmaB. The sigB coding region is preceded by four open reading frames (ORFs): dpsA, orfA, rsbB and rsbA in sequential order. RNA analyses revealed that rsbB, rsbA and sigB constitute an operon (sigB operon). Transcripts were produced constitutively from a promoter (sigBp2) upstream of the rsbB coding region, contributing to the basal level expression of sigmaB protein. An inducible promoter (sigBp1) resembling the catB promoter (catBp) was located between the rsbA and sigB coding regions. Transcripts from sigBp1 dramatically increased as cells differentiated on solid media, at the stationary phase in liquid media or by osmotic stresses similar to the behaviour of catBp transcripts. Both catBp and sigBp1 promoters were recognized specifically by sigmaB-containing RNA polymerase in vitro. Disruption of the sigB gene abolished not only the differentiation-associated expression but also the osmotic induction of the catB gene, indicating that catBp is under the control of sigmaB. The sigB mutant exhibited a similar phenotype to the catB mutant, being sensitive to hyperosmolarity, blocked in forming aerial mycelium and with skewed antibiotic production. Therefore, we conclude that sigmaB ensures the proper differentiation and osmoprotection of S. coelicolor cells, primarily via regulation of the expression of catalase B.
在天蓝色链霉菌A3(2)中分离并鉴定了一个编码替代σ因子σB的基因(sigB)。它编码一个由281个氨基酸组成的多肽(31546 Da),与枯草芽孢杆菌的σB高度同源。sigB编码区之前依次有四个开放阅读框(ORF):dpsA、orfA、rsbB和rsbA。RNA分析表明,rsbB、rsbA和sigB构成一个操纵子(sigB操纵子)。转录本由rsbB编码区上游的一个启动子(sigBp2)组成型产生,有助于σB蛋白的基础水平表达。一个类似于catB启动子(catBp)的可诱导启动子(sigBp1)位于rsbA和sigB编码区之间。当细胞在固体培养基上分化、在液体培养基的稳定期或受到与catBp转录本行为相似的渗透胁迫时,来自sigBp1的转录本显著增加。catBp和sigBp1启动子在体外都能被含σB的RNA聚合酶特异性识别。sigB基因的破坏不仅消除了与分化相关的表达,也消除了catB基因的渗透诱导,表明catBp受σB的控制。sigB突变体表现出与catB突变体相似的表型,对高渗敏感,气生菌丝形成受阻,抗生素产生偏向。因此,我们得出结论,σB主要通过调节过氧化氢酶B的表达来确保天蓝色链霉菌细胞的正常分化和渗透保护。