Ramírez J, Ramírez O, Saldaña C, Coria R, Peña A
Departamento de Genética Molecular, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, 04510 México D. F., México.
J Bacteriol. 1998 Nov;180(22):5860-5. doi: 10.1128/JB.180.22.5860-5865.1998.
The KHA1 gene corresponding to the open reading frame YJL094c (2.62 kb) encoding a putative K+/H+ antiporter (873 amino acids) in Saccharomyces cerevisiae was disrupted by homologous recombination. The core protein is similar to the putative Na+/H+ antiporters from Enterococcus hirae (NAPA gene) and Lactococcus lactis (LLUPP gene) and the putative K+/H+ exchanger from Escherichia coli (KEFC gene). Disruption of the KHA1 gene resulted in an increased K+ accumulation and net influx without a significant difference in efflux, as well as an increased growth rate, smaller cells, and twice the cell yield per glucose used. Flow cytometry analysis showed an increase of the DNA duplication rate in the mutant. Kinetic studies of 86Rb+ uptake showed the same saturable system for wild-type and disruptant strains. Mutant cells also produced a greater acidification of the medium coincident with an internal pH alkalinization and showed a higher oxygen consumption velocity. We speculate that higher K+ accumulation and increased osmotic pressure accelerate the cell cycle and metabolic activity.
通过同源重组破坏了酿酒酵母中与开放阅读框YJL094c(2.62 kb)对应的KHA1基因,该开放阅读框编码一种假定的K⁺/H⁺逆向转运蛋白(873个氨基酸)。核心蛋白与来自平肠球菌的假定Na⁺/H⁺逆向转运蛋白(NAPA基因)、乳酸乳球菌的假定Na⁺/H⁺逆向转运蛋白(LLUPP基因)以及大肠杆菌的假定K⁺/H⁺交换蛋白(KEFC基因)相似。KHA1基因的破坏导致K⁺积累增加和净流入增加,而流出没有显著差异,同时生长速率增加、细胞变小,每消耗的葡萄糖产生的细胞产量增加两倍。流式细胞术分析表明突变体中DNA复制率增加。对⁸⁶Rb⁺摄取的动力学研究表明野生型和破坏菌株具有相同的可饱和系统。突变细胞还使培养基产生更大程度的酸化,同时细胞内pH值碱化,并显示出更高的耗氧速度。我们推测较高的K⁺积累和渗透压增加会加速细胞周期和代谢活性。