Homuth G, Mogk A, Schumann W
Institute of Genetics, University of Bayreuth, D-95440 Bayreuth, Germany.
Mol Microbiol. 1999 Jun;32(6):1183-97. doi: 10.1046/j.1365-2958.1999.01428.x.
The heptacistronic dnaK heat shock operon of Bacillus subtilis consists of the genes hrcA, grpE, dnaK, dnaJ, orf35, orf28 and orf50. It is controlled by the CIRCE/HrcA operator/repressor system and specifies three primary transcripts, two of which are processed into three different products. We have analysed the regulatory consequences of this complex transcriptional organization in detail. First, the seven genes were heat induced to different extents at the mRNA level and can be classified into three groups by their induction factors. This differential induction was also reflected at the protein level. Secondly, the cellular amounts of the proteins HrcA, DnaK and DnaJ in B. subtilis differed drastically both under non-heat shock conditions and after thermal upshock. Thirdly, Northern blot analyses demonstrated that an mRNA-processing reaction generating products of differential stabilities plays an essential role during the regulation of gene expression. A crucial factor determining the low stability of two transcripts is the presence of the CIRCE element at their 5' ends. We demonstrate that CIRCE leads to the destabilization of mRNAs, but only if it is located in the immediate vicinity of a Shine-Dalgarno sequence. These results show that B. subtilis is using various, especially post-transcriptional, regulatory mechanisms to fine tune the expression of the individual genes of the heptacistronic dnaK operon.
枯草芽孢杆菌的七顺反子dnaK热休克操纵子由hrcA、grpE、dnaK、dnaJ、orf35、orf28和orf50基因组成。它受CIRCE/HrcA操纵子/阻遏物系统控制,并指定三种主要转录本,其中两种被加工成三种不同的产物。我们详细分析了这种复杂转录组织的调控后果。首先,这七个基因在mRNA水平上受热诱导的程度不同,根据其诱导因子可分为三组。这种差异诱导在蛋白质水平上也有体现。其次,枯草芽孢杆菌中HrcA、DnaK和DnaJ蛋白的细胞含量在非热休克条件下和热激后都有很大差异。第三,Northern印迹分析表明,产生稳定性不同的产物的mRNA加工反应在基因表达调控过程中起重要作用。决定两种转录本低稳定性的关键因素是其5'端存在CIRCE元件。我们证明CIRCE会导致mRNA不稳定,但前提是它位于Shine-Dalgarno序列的紧邻区域。这些结果表明,枯草芽孢杆菌正在利用各种调控机制,尤其是转录后调控机制,来微调七顺反子dnaK操纵子中各个基因的表达。