Price C W, Doi R H
Mol Gen Genet. 1985;201(1):88-95. doi: 10.1007/BF00397991.
We have mapped the chromosomal locus of rpoD, which encodes the major sigma factor of Bacillus subtilis RNA polymerase. The rpoD locus lay between aroD and lys, tightly linked to dnaE and inseparable from crsA. Marker order in this region was acf-aroD-dnaE-rpoD(crsA)-spoOG-lys. By transformation using cloned donor DNA from the rpoD region, we identified the gene immediately upstream of rpoD as dnaE, which coded for a 62,000 dalton protein essential for DNA replication. Both dnaE and rpoD were transcribed in the same direction, counterclockwise on the chromosome. The gene functions and organization in the rpoD region are thus similar to those of the E. coli sigma operon. We also used transformation to identify crsA47 as a mutation within the sigma coding region itself. The crsA alteration of sigma renders the sporulation process insensitive to glucose catabolite repression, and also restores sporulation ability to strains carrying early-blocked spoOE, spoOF, and spoOK mutations. Thus the major sigma factor and these spoO gene products directly or indirectly affect the same cellular function.
我们已绘制出rpoD的染色体定位,rpoD编码枯草芽孢杆菌RNA聚合酶的主要σ因子。rpoD基因座位于aroD和lys之间,与dnaE紧密连锁且与crsA不可分离。该区域的标记顺序为acf - aroD - dnaE - rpoD(crsA)- spoOG - lys。通过使用来自rpoD区域的克隆供体DNA进行转化,我们确定rpoD上游紧邻的基因为dnaE,它编码一种对DNA复制至关重要的62,000道尔顿蛋白质。dnaE和rpoD均沿相同方向转录,即在染色体上呈逆时针方向。因此,rpoD区域的基因功能和组织与大肠杆菌的σ操纵子相似。我们还利用转化确定crsA47是σ编码区域内的一个突变。σ的crsA改变使芽孢形成过程对葡萄糖分解代谢物阻遏不敏感,并且还恢复了携带早期阻断的spoOE、spoOF和spoOK突变的菌株的芽孢形成能力。因此,主要的σ因子和这些spoO基因产物直接或间接影响相同的细胞功能。