AUTHOR=Gu Huawei , Yang Yingyin , Wang Meng , Chen Shuyi , Wang Haiying , Li Shan , Ma Yi , Wang Jufang 

TITLE=Novel Cysteine Desulfidase CdsB Involved in Releasing Cysteine Repression of Toxin Synthesis in Clostridium difficile

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=7

YEAR=2018

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2017.00531

DOI=10.3389/fcimb.2017.00531

ISSN=2235-2988

ABSTRACT=<p><italic>Clostridium difficile</italic>, a major cause of nosocomial diarrhea and pseudomembranous colitis, still poses serious health-care challenges. The expression of its two main virulence factors, TcdA and TcdB, is reportedly repressed by cysteine, but molecular mechanism remains unclear. The cysteine desulfidase CdsB affects the virulence and infection progresses of some bacteria. The <italic>C. difficile</italic> strain 630 genome encodes a homolog of CdsB, and in the present study, we analyzed its role in <italic>C. difficile</italic> 630Δ<italic>erm</italic> by constructing an isogenic ClosTron-based <italic>cdsB</italic> mutant. When <italic>C. difficile</italic> was cultured in TY broth supplemented with cysteine, the <italic>cdsB</italic> gene was rapidly induced during the exponential growth phase. The inactivation of <italic>cdsB</italic> not only affected the resistance of <italic>C. difficile</italic> to cysteine, but also altered the expression levels of intracellular cysteine-degrading enzymes and the production of hydrogen sulfide. This suggests that <italic>C. difficile</italic> CdsB is a major inducible cysteine-degrading enzyme. The inactivation of the <italic>cdsB</italic> gene in <italic>C. difficile</italic> also removed the cysteine-dependent repression of toxin production, but failed to remove the Na<sub>2</sub>S-dependent repression, which supports that the cysteine-dependent repression of toxin production is probably attributable to the accumulation of cysteine by-products. We also mapped a δ<sup>54</sup> (SigL)-dependent promoter upstream from the <italic>cdsB</italic> gene, and <italic>cdsB</italic> expression was not induced in response to cysteine in the <italic>cdsR</italic>::<italic>ermB</italic> or <italic>sigL</italic>::<italic>ermB</italic> strain. Using a reporter gene fusion analysis, we identified the necessary promoter sequence for cysteine-dependent <italic>cdsB</italic> expression. Taken together, these results indicate that CdsB is a key inducible cysteine desulfidase in <italic>C. difficile</italic> which is regulated by δ<sup>54</sup> and CdsR in response to cysteine and that cysteine-dependent regulation of toxin production is closely associated with cysteine degradation.</p>