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应激反应σ因子σ(H)控制天蓝色链霉菌A3(2)中ssgB的表达,ssgB是芽孢形成特异性细胞分裂基因ssgA的同源物。

The stress-response sigma factor sigma(H) controls the expression of ssgB, a homologue of the sporulation-specific cell division gene ssgA, in Streptomyces coelicolor A3(2).

作者信息

Kormanec J, Sevcikova B

机构信息

Institute of Molecular Biology, Slovak Academy of Sciences, Dubravska cesta 21, 842 51 Bratislava, Slovak Republic.

出版信息

Mol Genet Genomics. 2002 Jun;267(4):536-43. doi: 10.1007/s00438-002-0687-0. Epub 2002 May 18.

Abstract

By using a previously established method for the identification of promoters recognized by a particular sigma factor of RNA polymerase, we identified a promoter in Streptomyces coelicolor A3(2) that is recognized by a heterologous RNA polymerase containing the late sporulation-specific sigma factor sigma(F). The promoter directed the expression of a gene named ssgB, which is related to the sporulation-specific cell division gene ssgA. These genes, together with three others, constitute a new family of paralogous genes specific for Streptomyces. S1-nuclease mapping using RNA prepared from an Escherichia coli strain that expresses ssgB under the control of sigma(F), and from S. coelicolor A3(2) at various developmental stages, identified identical transcription start points in both strains, corresponding to the promoter ssgBp. The promoter is developmentally regulated in S. coelicolor: it is induced at the time of aerial mycelium formation and is most active during sporulation. However, the level of the ssgB transcript was unaffected in a sigF mutant of S. coelicolor A3(2). Interestingly, the level of the transcript was substantially reduced in an S. coelicolor strain that was mutant for the sigH gene, which encodes a stress-response sigma factor (sigma(H)) that is essential for sporulation in S. coelicolor A3(2). This dependence of ssgB upon sigH was confirmed by an in vitro transcription assay, in which sigma(H), in the presence of the S. coelicolor core RNA polymerase, was able to recognize ssgBp. These results suggest that the S. coelicolor ssgB gene is under the control of the stress-response sigma(H). Transcription of ssgB was investigated in S. coelicolor A3(2) mutants with lesions in each of six known early whi genes required for sporulation septation. Expression of ssgB was deregulated in three of the mutants ( whiA, whiI, and whiJ). Based on these data, it is proposed that the ssgB gene product plays a role in the developmental process in S. coelicolor A3(2).

摘要

通过使用先前建立的用于鉴定由RNA聚合酶的特定σ因子识别的启动子的方法,我们在天蓝色链霉菌A3(2)中鉴定出一个启动子,该启动子可被含有晚期孢子形成特异性σ因子σ(F)的异源RNA聚合酶识别。该启动子指导一个名为ssgB的基因的表达,该基因与孢子形成特异性细胞分裂基因ssgA相关。这些基因与其他三个基因一起,构成了一个新的链霉菌特异性旁系同源基因家族。使用从在σ(F)控制下表达ssgB的大肠杆菌菌株以及在不同发育阶段的天蓝色链霉菌A3(2)制备的RNA进行S1核酸酶图谱分析,确定了两种菌株中相同的转录起始点,对应于启动子ssgBp。该启动子在天蓝色链霉菌中受到发育调控:它在气生菌丝形成时被诱导,并且在孢子形成期间最活跃。然而,在天蓝色链霉菌A3(2)的sigF突变体中,ssgB转录本的水平未受影响。有趣的是,在编码应激反应σ因子σ(H)(天蓝色链霉菌A3(2)孢子形成所必需)的sigH基因突变的天蓝色链霉菌菌株中,转录本水平大幅降低。通过体外转录试验证实了ssgB对σ(H)的这种依赖性,在该试验中,σ(H)在天蓝色链霉菌核心RNA聚合酶存在的情况下能够识别ssgBp。这些结果表明,天蓝色链霉菌的ssgB基因受应激反应σ(H)的控制。在参与孢子形成隔膜的六个已知早期whi基因中每个基因都有损伤的天蓝色链霉菌A3(2)突变体中研究了ssgB的转录。在三个突变体(whiA、whiI和whiJ)中,ssgB的表达失调。基于这些数据,有人提出ssgB基因产物在天蓝色链霉菌A3(2)的发育过程中起作用。

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