Kim Tae-Hyun, Park Joon-Sung, Kim Hyung-Joon, Kim Younhee, Kim Pil, Lee Heung-Shick
Graduate School of Biotechnology, Korea University, Anam-Dong, Sungbuk-Ku, Seoul 136-701, Republic of Korea.
Biochem Biophys Res Commun. 2005 Nov 25;337(3):757-64. doi: 10.1016/j.bbrc.2005.09.115. Epub 2005 Sep 28.
In this study, we have analyzed an ORF from Corynebacterium glutamicum, which codes for a homologue of the Streptomyces coelicolor WhiB-family of proteins known to be involved in sporulation. This ORF encoded a putative protein which harbors a helix-turn-helix DNA-binding motif and a probable redox-sensing motif, and has been designated whcE. We constructed a whcE mutant strain and analyzed the strain under a variety of growth conditions. This mutant strain exhibited a prolonged lag phase and earlier death within the stationary phase, suggesting that the relevant gene may play a role in both growth adaptation and stress responses. Further analysis determined that the mutant strain was not only sensitive with regard to survival under heat stress, but was also markedly susceptible to thiol-specific oxidant diamide and redox cycling compounds, including menadione and plumbagin. The mutant strain also exhibited reductions in thioredoxin reductase activity, which indicates that the trxB gene encoding thioredoxin reductase is under the control of WhcE. Expression of whcE was stimulated during the stationary phase of cell growth and could be modulated by diamide. We also delineated the relationship between whcE and the sigH gene, which is located downstream of whcE, and has been shown to be involved in heat stress responses, via the encoding of an ECF sigma factor. In a sigH mutant strain, the whcE gene was no longer expressed, thereby suggesting that the sigmaH sigma factor is involved in whcE expression. Our results suggest that WhcE functions as a transcription factor which can activate the trxB gene, as well as other genes, possibly by sensing redox changes during the metabolic downshifting of cells from exponential growth to the stationary phase, whereas sigmaH appears to function as the sigma factor for these genes, including whcE.
在本研究中,我们分析了谷氨酸棒杆菌的一个开放阅读框(ORF),其编码天蓝色链霉菌WhiB家族蛋白的同源物,已知该家族蛋白参与孢子形成。这个开放阅读框编码一种假定的蛋白质,它含有一个螺旋-转角-螺旋DNA结合基序和一个可能的氧化还原感应基序,并被命名为whcE。我们构建了一个whcE突变株,并在多种生长条件下对该菌株进行了分析。该突变株表现出延长的延滞期和在稳定期更早的死亡,这表明相关基因可能在生长适应和应激反应中都发挥作用。进一步分析确定,该突变株不仅在热应激下生存敏感,而且对硫醇特异性氧化剂二酰胺和氧化还原循环化合物(包括甲萘醌和白花丹素)也明显敏感。该突变株还表现出硫氧还蛋白还原酶活性降低,这表明编码硫氧还蛋白还原酶的trxB基因受WhcE的控制。whcE的表达在细胞生长的稳定期受到刺激,并且可以被二酰胺调节。我们还描绘了whcE与sigH基因之间的关系,sigH基因位于whcE下游,已被证明通过编码一种额外的细胞质因子(ECF)σ因子参与热应激反应。在sigH突变株中,whcE基因不再表达,从而表明σH σ因子参与whcE的表达。我们的结果表明,WhcE作为一种转录因子,可能通过感知细胞从指数生长向稳定期代谢下降过程中的氧化还原变化来激活trxB基因以及其他基因,而σH似乎作为这些基因(包括whcE)的σ因子发挥作用。