Quan John A, Schneider Barbara L, Paulsen Ian T, Yamada Mamoru, Kredich Nicholas M, Saier Milton H
Department of Biology, University of California at San Diego, La Jolla, CA 92093-0116, USA1.
Department of Biochemistry, Yamaguchi University School of Medicine, Ube, Yamaguchi, Japan 7552.
Microbiology (Reading). 2002 Jan;148(Pt 1):123-131. doi: 10.1099/00221287-148-1-123.
Different pleiotropic transcriptional regulators are known to function in the coordination of regulons concerned with carbon, nitrogen, sulfur, phosphorus and iron metabolism, but how expression profiles of these different regulons are coordinated with each other is not known. The basis for the effects of cysB mutations on carbon utilization in Escherichia coli and Salmonella typhimurium was examined. cysB mutations affected the utilization of some carbon sources more than others and these effects could be partially, but not completely, reversed by the inclusion of cysteine or djenkolate in the growth medium. Assays of transport systems and enzymes concerned with glucitol and alanine utilization showed that these activities were depressed in cysB mutants relative to isogenic wild-type strains, and cysteine or djenkolate present in the growth media partially restored these activities. Using transcriptional fusions to the fdo (formate dehydrogenase) and gut (glucitol) operons, it was shown that decreased expression resulted from defects at the transcriptional level. Furthermore, the effects of loss of CysB were much less pronounced under conditions of catabolite repression than in the absence of a catabolite-repressing carbon source, and cAMP largely reversed the effect of the loss of CysB. Comparable effects were seen for E. coli lacZ gene expression under the control of its own native promoter, and sulfur limitation in a cysB mutant depressed net cAMP production in a cAMP phosphodiesterase mutant. Adenylate cyclase thus appears to be responsive to sulfur deprivation. These observations may have physiological significance allowing carbon and sulfur regulon coordination during the growth of enteric bacteria in response to nutrient availability.
已知不同的多效转录调节因子在与碳、氮、硫、磷和铁代谢相关的调节子协调中发挥作用,但这些不同调节子的表达谱如何相互协调尚不清楚。研究了cysB突变对大肠杆菌和鼠伤寒沙门氏菌碳利用的影响基础。cysB突变对某些碳源利用的影响大于其他碳源,并且通过在生长培养基中添加半胱氨酸或木豆叶可以部分但不完全逆转这些影响。对与葡糖醇和丙氨酸利用相关的转运系统和酶的测定表明,相对于同基因野生型菌株,这些活性在cysB突变体中受到抑制,生长培养基中存在的半胱氨酸或木豆叶部分恢复了这些活性。利用与fdo(甲酸脱氢酶)和gut(葡糖醇)操纵子的转录融合,表明表达降低是由转录水平的缺陷导致的。此外,在分解代谢物阻遏条件下,CysB缺失的影响比在没有分解代谢物阻遏碳源的情况下要小得多,并且cAMP在很大程度上逆转了CysB缺失的影响。在其自身天然启动子控制下的大肠杆菌lacZ基因表达也观察到了类似的效果,并且cysB突变体中的硫限制降低了cAMP磷酸二酯酶突变体中的净cAMP产生。因此,腺苷酸环化酶似乎对硫缺乏有反应。这些观察结果可能具有生理意义,允许肠道细菌在生长过程中根据营养可用性协调碳和硫调节子。