Mueller J P, Taber H W
Department of Microbiology and Immunology, Albany Medical College, New York 12208.
J Bacteriol. 1989 Sep;171(9):4979-86. doi: 10.1128/jb.171.9.4979-4986.1989.
Mutations that define the ctaA gene of Bacillus subtilis block cytochrome aa3 formation and sporulation. We have recently described the isolation and initial characterization of the ctaA locus. Analysis of in vivo mRNA transcripts by RNase protection experiments located the 5' and 3' termini of the ctaA transcript, confirming a monocistronic structure. By using a nuclease protection assay, an increase in the abundance of steady-state ctaA mRNA was observed during the initiation of sporulation, followed by a decrease during subsequent stages. Transcripts originating from the ctaA gene were most abundant 2.0 h after the end of exponential growth. This pattern of ctaA mRNA accumulation was confirmed by coupling the transcription of the ctaA gene to lacZ in an integrative plasmid vector. Expression of ctaA was not repressed by glucose and was independent of the spoOA and spoOH (sigH) gene products. Postexponential expression was found to be dependent on the product of the strC gene. The expression of ctaA appears to be regulated in a growth stage-specific manner. The transcriptional start site, identified by high-resolution S1 nuclease protection experiments, was preceded by a single sigma A-dependent promoter sequence.
定义枯草芽孢杆菌ctaA基因的突变会阻断细胞色素aa3的形成和芽孢形成。我们最近描述了ctaA基因座的分离和初步特征。通过核糖核酸酶保护实验对体内mRNA转录本进行分析,确定了ctaA转录本的5'和3'末端,证实其为单顺反子结构。通过核酸酶保护试验,在芽孢形成起始阶段观察到稳态ctaA mRNA丰度增加,随后在后续阶段下降。源自ctaA基因的转录本在指数生长结束后2.0小时最为丰富。通过在整合质粒载体中将ctaA基因的转录与lacZ偶联,证实了ctaA mRNA的这种积累模式。ctaA的表达不受葡萄糖抑制,且独立于spoOA和spoOH(sigH)基因产物。发现指数期后表达依赖于strC基因的产物。ctaA的表达似乎以生长阶段特异性方式受到调节。通过高分辨率S1核酸酶保护实验确定的转录起始位点之前有一个单一的依赖于sigma A的启动子序列。