Robertson J B, Gocht M, Marahiel M A, Zuber P
Department of Botany and Microbiology, Oklahoma State University, Stillwater 74078.
Proc Natl Acad Sci U S A. 1989 Nov;86(21):8457-61. doi: 10.1073/pnas.86.21.8457.
The abrB gene of Bacillus subtilis is believed to encode a repressor that controls the expression of genes involved in starvation-induced processes such as sporulation and the production of antibiotics and degradative enzymes. Two such genes, spoVG, a sporulation gene of B. subtilis, and tycA, which encodes tyrocidine synthetase I of the tyrocidine biosynthetic pathway in Bacillus brevis, are negatively regulated by abrB in B. subtilis. To examine the role of abrB in the repression of gene transcription, the AbrB protein was purified and then tested for its ability to bind to spoVG and tycA promoter DNA. In a gel mobility shift experiment, AbrB was found to bind to a DNA fragment containing the sequence from -95 to +61 of spoVG. AbrB protein exhibited reduced affinity for DNA of two mutant forms of the spoVG promoter that had been shown to be insensitive to abrB-dependent repression in vivo. These studies showed that an upstream A + T-rich sequence from -37 to -95 was required for optimal AbrB binding. AbrB protein was also observed to bind to the tycA gene within a region between the transcription start site and the tycA coding sequence as well as to a region containing the putative tycA promoter. These findings reinforce the hypothesis that AbrB represses gene expression through its direct interaction with the transcription initiation regions of genes under its control.
枯草芽孢杆菌的abrB基因被认为编码一种阻遏蛋白,该蛋白控制参与饥饿诱导过程的基因表达,如芽孢形成、抗生素生产和降解酶的产生。枯草芽孢杆菌中的两个这样的基因,即spoVG(枯草芽孢杆菌的一个芽孢形成基因)和tycA(编码短短芽孢杆菌中短杆菌肽生物合成途径的短杆菌肽合成酶I),在枯草芽孢杆菌中受到abrB的负调控。为了研究abrB在基因转录抑制中的作用,纯化了AbrB蛋白,然后测试其与spoVG和tycA启动子DNA结合的能力。在凝胶迁移率变动实验中,发现AbrB与一个包含spoVG从-95到+61序列的DNA片段结合。AbrB蛋白对spoVG启动子的两种突变形式的DNA亲和力降低,这两种突变形式在体内已被证明对abrB依赖性抑制不敏感。这些研究表明,-37至-95的上游富含A+T的序列是AbrB最佳结合所必需的。还观察到AbrB蛋白在转录起始位点和tycA编码序列之间的区域以及包含假定tycA启动子的区域内与tycA基因结合。这些发现强化了这样一种假设,即AbrB通过与受其控制的基因的转录起始区域直接相互作用来抑制基因表达。