A. A. Kharkevich Institute for Information Transmission Problems, RAS, Bolshoy Karetny per. 19, Moscow, 127051, Russia.
N. I. Vavilov Institute of General Genetics, RAS, ul. Gubkina 3, Moscow, 119991, Russia.
Sci Rep. 2018 Feb 16;8(1):3177. doi: 10.1038/s41598-018-21534-3.
Comparative genomics analysis of conserved gene cassettes demonstrated resemblance between a recently described cassette of genes involved in sulphoquinovose degradation in Escherichia coli K-12 MG1655 and a Bacilli cassette linked with lactose degradation. Six genes from both cassettes had similar functions related to carbohydrate metabolism, namely, hydrolase, aldolase, kinase, isomerase, transporter, and transcription factor. The Escherichia coli sulphoglycolysis cassette was thus predicted to be associated with lactose degradation. This prediction was confirmed experimentally: expression of genes coding for aldolase (yihT), isomerase (yihS), and kinase (yihV) was dramatically increased during growth on lactose. These genes were previously shown to be activated during growth on sulphoquinovose, so our observation may indicate multi-functional capabilities of the respective proteins. Transcription starts for yihT, yihV and yihW were mapped in silico, in vitro and in vivo. Out of three promoters for yihT, one was active only during growth on lactose. We further showed that switches in yihT transcription are controlled by YihW, a DeoR-family transcription factor in the Escherichia coli cassette. YihW acted as a carbon source-dependent dual regulator involved in sustaining the baseline growth in the absence of lac-operon, with function either complementary, or opposite to a global regulator of carbohydrate metabolism, cAMP-CRP.
比较基因组学分析保守基因盒表明,在大肠杆菌 K-12 MG1655 中最近描述的参与硫酸奎诺糖降解的基因盒与与乳糖降解相关的芽孢杆菌盒之间存在相似性。这两个盒的六个基因具有相似的碳水化合物代谢相关功能,即水解酶、醛缩酶、激酶、异构酶、转运蛋白和转录因子。因此,预测大肠杆菌硫酸盐降解盒与乳糖降解有关。这一预测得到了实验的证实:在乳糖上生长时,编码醛缩酶(yihT)、异构酶(yihS)和激酶(yihV)的基因表达显著增加。这些基因以前在生长于硫酸奎诺糖时被激活,因此我们的观察结果可能表明各自蛋白质的多功能能力。yihT、yihV 和 yihW 的转录起始在计算机上、体外和体内进行了映射。在 yihT 的三个启动子中,只有一个在乳糖上生长时才活跃。我们进一步表明,yihT 转录的开关由 YihW 控制,YihW 是大肠杆菌盒中的 DeoR 家族转录因子。YihW 作为碳源依赖性双重调节剂,参与在 lac 操纵子缺失的情况下维持基线生长,其功能要么与碳水化合物代谢的全局调节剂 cAMP-CRP 互补,要么相反。