Department of Biomedicinal Science and Biotechnology, Paichai University, Daejeon 302-735, Republic of Korea.
J Microbiol Biotechnol. 2013 Dec;23(12):1683-90. doi: 10.4014/jmb.1310.10031.
The cyclic AMP receptor protein (CRP) homolog, GlxR, controls the expression of several genes involved in the regulation of diverse physiological processes in Corynebacterium glutamicum. In silico analysis has revealed the presence of glxR binding sites upstream of genes adhA, ald, and ptsG, encoding glucose-specific phosphotransferase system protein, alcohol dehydrogenase (ADH), and acetaldehyde dehydrogenase (ALDH), respectively. However, the involvement of the GlxR-cAMP complex on the expression of these genes has been explored only in vitro. In this study, the expressions of ptsG, adhA, and ald were analyzed in detail using an adenylate cyclase gene (cyaB) deletion mutant and glxR deletion mutant. The specific activities of ADH and ALDH were increased in both the mutants in glucose and glucose plus ethanol media, in contrast to the wild type. In accordance, the promoter activities of adhA and ald were derepressed in the cyaB mutant, indicating that glxR acts as a repressor of adhA. Similarly, both the mutants exhibited derepression of ptsG regardless of the carbon source. These results confirm the involvement of GlxR on the expression of important carbon metabolic genes; adhA, ald, and ptsG.
环腺苷酸受体蛋白 (CRP) 同源物 GlxR 控制着几种基因的表达,这些基因参与了谷氨酸棒杆菌中多种生理过程的调节。计算机分析表明,adhA、ald 和 ptsG 基因上游存在 glxR 结合位点,分别编码葡萄糖特异性磷酸转移酶系统蛋白、醇脱氢酶 (ADH) 和乙醛脱氢酶 (ALDH)。然而,GlxR-cAMP 复合物对这些基因表达的影响仅在体外进行了探索。在这项研究中,使用腺嘌呤环化酶基因 (cyaB) 缺失突变体和 glxR 缺失突变体详细分析了 ptsG、adhA 和 ald 的表达。与野生型相比,在葡萄糖和葡萄糖加乙醇培养基中,两种突变体的 ADH 和 ALDH 比活性都增加。相应地,在 cyaB 突变体中,adhA 和 ald 的启动子活性被去阻遏,表明 GlxR 作为 adhA 的阻遏物发挥作用。同样,无论碳源如何,两种突变体都表现出 ptsG 的去阻遏。这些结果证实了 GlxR 参与了重要的碳代谢基因的表达;adhA、ald 和 ptsG。