Bryan E M, Beall B W, Moran C P
Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
J Bacteriol. 1996 Aug;178(16):4778-86. doi: 10.1128/jb.178.16.4778-4786.1996.
To identify genes expressed at intermediate stages of Bacillus subtilis sporulation, we screened for sigma E-dependent promoters. One promoter that we found drives expression of an operon consisting of at least five open reading frames (ORFs). The predicted products of the first three ORFs are very homologous to enzymes involved in fatty acid metabolism, including acetyl coenzyme A (acetyl-CoA) acetyltransferase (thiolase), 3-hydroxybutyryl-CoA dehydrogenase, and acyl-CoA dehydrogenase, respectively. We showed that the fourth ORF encoded a third isozyme of citrate synthase in B. subtilis. Genetic evidence and primer extension results showed that transcription of this operon is directed by the mother cell compartment-specific sigma factor, sigma E, and so the operon was named mmg (for mother cell metabolic genes). Furthermore, we found that a sequence (mmgO) with homology to a catabolite-responsive element mediates glucose repression of mmg promoter activity during sporulation and that this repression was lost in a ccpA mutant.
为了鉴定在枯草芽孢杆菌芽孢形成中间阶段表达的基因,我们筛选了依赖于σE的启动子。我们发现的一个启动子驱动一个由至少五个开放阅读框(ORF)组成的操纵子的表达。前三个ORF的预测产物分别与参与脂肪酸代谢的酶高度同源,包括乙酰辅酶A(乙酰-CoA)乙酰转移酶(硫解酶)、3-羟基丁酰-CoA脱氢酶和酰基辅酶A脱氢酶。我们证明第四个ORF编码枯草芽孢杆菌柠檬酸合酶的第三种同工酶。遗传证据和引物延伸结果表明,该操纵子的转录由母细胞区室特异性σ因子σE指导,因此该操纵子被命名为mmg(母细胞代谢基因)。此外,我们发现一个与分解代谢物反应元件具有同源性的序列(mmgO)介导了芽孢形成过程中mmg启动子活性的葡萄糖抑制作用,并且这种抑制作用在ccpA突变体中消失。