Viala J, Rapoport G, Mazodier P
Unité de Biochimie Microbienne, CNRS URA 2172, Institut Pasteur, 25 rue du Docteur Roux, 75724 Paris Cedex 15, France.
Mol Microbiol. 2000 Nov;38(3):602-12. doi: 10.1046/j.1365-2958.2000.02155.x.
The clpP genes are widespread among living organisms and encode the proteolytic subunit of the Clp ATP-dependent protease. These genes are present in a single copy in most eubacteria. However, five clpP genes were identified in Streptomyces coelicolor. The clpP1 clpP2 operon was studied: mutations affected the growth cycle in various Streptomyces. Here, we report studies of the expression of the clpP3 clpP4 operon in Streptomyces lividans. The clpP3 operon was induced in a clpP1 mutant strain, and the regulation of expression was investigated in detail. The product of the putative regulator gene, downstream from clpP4, was purified. Gel migration shift assays and DNase I footprinting showed that this protein binds to the clpP3 promoter and recognizes a tandem 6 bp palindromic repeat (TCTGCC-3N-GGCAGA). In vivo, this DNA-binding protein, named PopR, acts as an activator of the clpP3 operon. Studies of popR expression indicate that the regulator is probably controlled at the post-transcriptional level.
clpP基因广泛存在于生物体内,编码Clp ATP依赖性蛋白酶的蛋白水解亚基。这些基因在大多数真细菌中以单拷贝形式存在。然而,在天蓝色链霉菌中鉴定出了五个clpP基因。对clpP1 clpP2操纵子进行了研究:突变影响了各种链霉菌的生长周期。在此,我们报告了对淡紫链霉菌中clpP3 clpP4操纵子表达的研究。clpP3操纵子在clpP1突变菌株中被诱导,并对其表达调控进行了详细研究。假定调控基因的产物在clpP4下游被纯化。凝胶迁移率变动分析和DNase I足迹分析表明,该蛋白与clpP3启动子结合,并识别一个串联的6 bp回文重复序列(TCTGCC-3N-GGCAGA)。在体内,这种名为PopR的DNA结合蛋白作为clpP3操纵子的激活剂发挥作用。对popR表达的研究表明,该调控因子可能在转录后水平受到控制。