University of Florida, Department of Microbiology and Cell Science, Gainesville, Florida 32611-0700, USA.
J Bacteriol. 2010 Dec;192(23):6251-60. doi: 10.1128/JB.00827-10. Epub 2010 Oct 8.
In this study, a DeoR/GlpR-type transcription factor was investigated for its potential role as a global regulator of sugar metabolism in haloarchaea, using Haloferax volcanii as a model organism. Common to a number of haloarchaea and Gram-positive bacterial species, the encoding glpR gene was chromosomally linked with genes of sugar metabolism. In H. volcanii, glpR was cotranscribed with the downstream phosphofructokinase (PFK; pfkB) gene, and the transcript levels of this glpR-pfkB operon were 10- to 20-fold higher when cells were grown on fructose or glucose than when they were grown on glycerol alone. GlpR was required for repression on glycerol based on significant increases in the levels of PFK (pfkB) transcript and enzyme activity detected upon deletion of glpR from the genome. Deletion of glpR also resulted in significant increases in both the activity and the transcript (kdgK1) levels of 2-keto-3-deoxy-D-gluconate kinase (KDGK), a key enzyme of haloarchaeal glucose metabolism, when cells were grown on glycerol, compared to the levels obtained for media with glucose. Promoter fusions to a β-galactosidase bgaH reporter revealed that transcription of glpR-pfkB and kdgK1 was modulated by carbon source and GlpR, consistent with quantitative reverse transcription-PCR (qRT-PCR) and enzyme activity assays. The results presented here provide genetic and biochemical evidence that GlpR controls both fructose and glucose metabolic enzymes through transcriptional repression of the glpR-pfkB operon and kdgK1 during growth on glycerol.
在这项研究中,我们研究了一种 DeoR/GlpR 型转录因子,以探讨其作为嗜盐古菌糖代谢全局调控因子的潜力,我们选择 Haloferax volcanii 作为模式生物。与许多嗜盐古菌和革兰氏阳性细菌物种一样,编码 glpR 的基因与糖代谢基因在染色体上相连。在 H. volcanii 中,glpR 与下游磷酸果糖激酶(PFK;pfkB)基因共转录,当细胞在果糖或葡萄糖上生长时,该 glpR-pfkB 操纵子的转录水平比在单独甘油上生长时高 10-20 倍。基于 glpR 缺失后 PFK(pfkB)转录本和酶活性的显著增加,glpR 对甘油的抑制是必需的。glpR 缺失也导致 2-酮-3-脱氧-D-葡萄糖酸激酶(KDGK)的活性和转录(kdgK1)水平显著增加,当细胞在甘油上生长时,与葡萄糖培养基中获得的水平相比,该酶是嗜盐古菌糖代谢的关键酶。与 β-半乳糖苷酶 bgaH 报告基因的启动子融合显示,glpR-pfkB 和 kdgK1 的转录受碳源和 GlpR 的调节,与定量逆转录-PCR(qRT-PCR)和酶活性测定结果一致。这里提供的遗传和生化证据表明,GlpR 通过转录抑制 glpR-pfkB 操纵子和 kdgK1 来控制果糖和葡萄糖代谢酶,在甘油生长过程中。