Mouncey N J, Kaplan S
Department of Microbiology and Molecular Genetics, The University of Texas Health Science Center Medical School, Houston, Texas 77030, USA.
J Bacteriol. 1998 Jun;180(11):2924-30. doi: 10.1128/JB.180.11.2924-2930.1998.
Under anaerobic-dark growth conditions, in the presence of the alternative electron acceptor dimethyl sulfoxide (DMSO) or trimethylamine N-oxide (TMAO), Rhodobacter sphaeroides 2.4.1(T) respires anaerobically using the molybdoenzyme DMSO reductase (DMSOR). Genes encoding DMSOR and associated proteins are encoded by genes of the dor locus. Previously, we demonstrated that the expression of DMSOR is regulated by both the oxygen status of the cell via the FnrL protein and by the presence of DMSO or TMAO, presumably through the DorS-DorR two-component sensor-regulator system. Here we further investigate expression of the dor genes through the use of transcriptional lacZ fusions to the dorS, dorR, and dorC promoters. The expression of dorC::lacZ was strongly induced by the absence of oxygen and presence of DMSO. In accordance with our previous findings of DMSOR activity, dorC::lacZ expression was reduced by up to one-third when cells were grown photosynthetically in the presence of DMSO with medium or high light, compared to the expression observed after anaerobic-dark growth. The induction of dorC::lacZ expression in the presence of DMSO was dependent on the DorS and DorR proteins. Expression of the dorS and dorR genes was also induced in the absence of oxygen. In an FnrL mutant, dorS::lacZ expression was not induced when oxygen tensions in the media were lowered, in contrast to what occurred in the wild-type strain. The expression of dorS::lacZ and dorR::lacZ was dependent on the DorS and DorR proteins themselves, suggesting the importance of autoregulation. These results demonstrate a cascade regulation of dor gene expression, where the expression of the regulatory proteins DorS and DorR governs the downstream regulation of the dorCBA operon encoding the structural proteins of DMSOR.
在厌氧黑暗生长条件下,当存在替代电子受体二甲基亚砜(DMSO)或三甲胺 N-氧化物(TMAO)时,球形红细菌 2.4.1(T)利用钼酶 DMSO 还原酶(DMSOR)进行厌氧呼吸。编码 DMSOR 和相关蛋白的基因由 dor 基因座的基因编码。此前,我们证明 DMSOR 的表达受细胞的氧状态通过 FnrL 蛋白调控,也受 DMSO 或 TMAO 的存在调控,推测是通过 DorS-DorR 双组分传感调节系统。在此,我们通过使用与 dorS、dorR 和 dorC 启动子的转录 lacZ 融合来进一步研究 dor 基因的表达。dorC::lacZ 的表达在无氧和存在 DMSO 时强烈诱导。与厌氧黑暗生长后观察到的表达相比,当细胞在存在 DMSO 的情况下进行光合生长且光照为中或高光时,dorC::lacZ 的表达降低了多达三分之一,这与我们之前关于 DMSOR 活性的发现一致。在存在 DMSO 时 dorC::lacZ 表达的诱导依赖于 DorS 和 DorR 蛋白。dorS 和 dorR 基因的表达在无氧时也被诱导。在 FnrL 突变体中,当培养基中的氧张力降低时,dorS::lacZ 的表达未被诱导,这与野生型菌株中的情况相反。dorS::lacZ 和 dorR::lacZ 的表达依赖于 DorS 和 DorR 蛋白本身,表明自动调节的重要性。这些结果证明了 dor 基因表达的级联调节,其中调节蛋白 DorS 和 DorR 的表达控制着编码 DMSOR 结构蛋白的 dorCBA 操纵子的下游调节。