Schulz A, Schwab S, Homuth G, Versteeg S, Schumann W
Institute of Genetics, University of Bayreuth, Germany.
J Bacteriol. 1997 May;179(10):3103-9. doi: 10.1128/jb.179.10.3103-3109.1997.
We show that the htpG gene of Bacillus subtilis is induced by heat, as has been reported for the Escherichia coli homolog. Analysis of different mutants revealed that the htpG gene belongs to class III heat shock genes in B. subtilis. An about 10-fold induction after thermal upshock was found at the levels of both transcription and translation, and this induction resulted from enhanced synthesis of mRNA. By primer extension, we identified one potential transcription start site immediately downstream of a putative sigmaA-dependent promoter which became activated after thermal upshift. Northern blot analysis revealed that htpG is part of a monocistronic transcriptional unit. An operon fusion where the complete region between htpG and its upstream gene was fused to the bgaB reporter gene accurately reflected htpG expression. Analysis of this fusion revealed that, in contrast to other class III heat shock genes, htpG was not induced by osmotic upshock, by ethanol, or by oxygen limitation, suggesting that it belongs to a subgroup within class III. Deletion of the region upstream of the putative promoter resulted in an enhanced basal level of htpG expression, but the 10-fold induction was retained, suggesting that the upstream sequences are involved in the regulation of expression in the absence of heat shock.
我们发现,枯草芽孢杆菌的htpG基因受热诱导,正如大肠杆菌的同源基因所报道的那样。对不同突变体的分析表明,htpG基因属于枯草芽孢杆菌中的III类热休克基因。热激后,转录和翻译水平均出现约10倍的诱导,这种诱导是由mRNA合成增强所致。通过引物延伸,我们在一个假定的依赖sigmaA的启动子下游立即鉴定出一个潜在的转录起始位点,该启动子在热激后被激活。Northern印迹分析表明,htpG是一个单顺反子转录单元的一部分。一个操纵子融合体,其中htpG与其上游基因之间的完整区域与bgaB报告基因融合,准确反映了htpG的表达。对该融合体的分析表明,与其他III类热休克基因不同,htpG不受渗透激、乙醇或氧限制的诱导,这表明它属于III类中的一个亚组。假定启动子上游区域的缺失导致htpG表达的基础水平升高,但10倍的诱导作用得以保留,这表明上游序列在无热休克时参与了表达调控。