Newell Katy V, Thomas Derek P, Brekasis Dimitris, Paget Mark S B
Department of Biochemistry, School of Life Sciences, University of Sussex, Falmer, Brighton BN1 9QG, UK.
Mol Microbiol. 2006 May;60(3):687-96. doi: 10.1111/j.1365-2958.2006.05116.x.
RbpA is an RNA polymerase-binding protein that occurs in the actinomycete family of bacteria and is regulated by the disulphide stress-response sigma factor, sigma(R), in Streptomyces coelicolor. Here we demonstrate that rbpA null mutants exhibit a slow-growth phenotype and are particularly sensitive to the transcription inhibitor rifampicin. Strikingly, transcription mapping experiments revealed that rbpA expression is induced upon exposure of S. coelicolor to rifampicin and that this, in part, involves an increase in the activity of sigma(R). In contrast, the ribosomal RNA operon promoter rrnDp3, which is recognized by the vegetative sigma factor sigma(HrdB), was strongly inhibited by rifampicin. Reconstitution of RNAP from an rbpA null mutant with purified RbpA revealed that RbpA stimulates transcription from rrnDp3, even in the presence of rifampicin. The data presented suggest that RbpA confers basal levels of rifampicin resistance and is a novel regulator of rRNA synthesis in S. coelicolor.
RbpA是一种RNA聚合酶结合蛋白,存在于放线菌科细菌中,在天蓝色链霉菌中受二硫键应激反应西格玛因子sigma(R)调控。在此我们证明,rbpA基因缺失突变体表现出生长缓慢的表型,并且对转录抑制剂利福平特别敏感。引人注目的是,转录图谱实验表明,天蓝色链霉菌暴露于利福平后rbpA表达被诱导,且这部分涉及sigma(R)活性的增加。相比之下,由营养型西格玛因子sigma(HrdB)识别的核糖体RNA操纵子启动子rrnDp3受到利福平的强烈抑制。用纯化的RbpA对rbpA基因缺失突变体的RNA聚合酶进行重组显示,即使在存在利福平的情况下,RbpA也能刺激rrnDp3的转录。所呈现的数据表明,RbpA赋予了利福平抗性的基础水平,并且是天蓝色链霉菌中rRNA合成的一种新型调节因子。