Cotter P A, Gunsalus R P
Department of Microbiology, University of California, Los Angeles.
J Bacteriol. 1989 Jul;171(7):3817-23. doi: 10.1128/jb.171.7.3817-3823.1989.
Escherichia coli can respire anaerobically using either trimethylamine-N-oxide (TMAO) or dimethyl sulfoxide (DMSO) as the terminal electron acceptor for oxidative phosphorylation. To determine whether the regulation of the dmsABC genes, which encode a membrane-associated TMAO/DMSO reductase, are transcriptionally controlled in response to the availability of alternate electron acceptors, we constructed an operon fusion between the dmsA gene, along with its associated regulatory region, and lacZ+. Expression of dmsA'-lacZ was stimulated 65-fold by anaerobiosis versus aerobiosis, while nitrate caused a 12-fold repression. Its expression, however, was unaffected by the presence of the alternate electron acceptors DMSO, TMAO, and fumarate. Anaerobic induction of dmsA'-lacZ was defective in an fnr mutant, thus establishing that Fnr is responsible for anaerobic activation of dmsABC. Repression of dmsA'-lacZ expression by nitrate was independent of oxygen and was shown to be mediated by the products of two genes, narL (frdR2) and narX. dmsA'-lacZ expression was also altered in chlD strains that are defective in molybdenum transport but not in chlA and chlE strains that are defective in molybdopterin cofactor biosynthesis, thus establishing that the molybdenum ion but not the ability to form a functional cofactor is required for regulation. Molybdenum was required both for complete induction of dmsA'-lacZ expression during anaerobic growth and for complete repression of dmsA'-lacZ by nitrate. Additionally, expression of dmsABC varied depending on the carbon source. Expression was highest when cells were grown on sorbitol.
大肠杆菌可以利用三甲胺 - N - 氧化物(TMAO)或二甲基亚砜(DMSO)作为氧化磷酸化的末端电子受体进行厌氧呼吸。为了确定编码膜相关TMAO / DMSO还原酶的dmsABC基因的调控是否响应替代电子受体的可用性而受到转录控制,我们构建了dmsA基因及其相关调控区域与lacZ +之间的操纵子融合体。与需氧相比,厌氧条件下dmsA'-lacZ的表达受到65倍的刺激,而硝酸盐则导致12倍的抑制。然而,其表达不受替代电子受体DMSO、TMAO和富马酸盐的影响。dmsA'-lacZ在fnr突变体中的厌氧诱导存在缺陷,从而确定Fnr负责dmsABC基因的厌氧激活。硝酸盐对dmsA'-lacZ表达的抑制与氧气无关,并且显示由两个基因narL(frdR2)和narX的产物介导。dmsA'-lacZ的表达在钼转运缺陷的chlD菌株中也发生了改变,但在钼蝶呤辅因子生物合成缺陷的chlA和chlE菌株中未发生改变,从而确定调控需要钼离子而不是形成功能性辅因子的能力。钼对于厌氧生长期间dmsA'-lacZ表达的完全诱导以及硝酸盐对dmsA'-lacZ的完全抑制都是必需的。此外,dmsABC的表达因碳源而异。当细胞在山梨醇上生长时,表达最高。