Lamb Teresa M, Mitchell Aaron P
Department of Microbiology and Institute of Cancer Research, Columbia University, New York, New York 10032, USA.
Mol Cell Biol. 2003 Jan;23(2):677-86. doi: 10.1128/MCB.23.2.677-686.2003.
Environmental pH changes have broad consequences for growth and differentiation. The best-understood eukaryotic pH response pathway acts through the zinc-finger transcription factor PacC of Aspergillus nidulans, which activates alkaline pH-induced genes directly. We show here that Saccharomyces cerevisiae Rim101p, the pH response regulator homologous to PacC, functions as a repressor in vivo. Chromatin immunoprecipitation assays show that Rim101p is associated in vivo with the promoters of seven Rim101p-repressed genes. A reporter gene containing deduced Rim101p binding sites is negatively regulated by Rim101p and is associated with Rim101p in vivo. Deletion mutations of the Rim101p repression targets NRG1 and SMP1 suppress rim101Delta mutant defects in ion tolerance, haploid invasive growth, and sporulation. Therefore, transcriptional repression is the main biological function of Rim101p. The Rim101p repression target Nrg1p is in turn required for repression of two alkaline pH-inducible genes, including the Na+ pump gene ENA1, which is required for ion tolerance. Thus, Nrg1p, a known transcriptional repressor, functions as an inhibitor of alkaline pH responses. Our findings stand in contrast to the well-characterized function of PacC as a direct activator of alkaline pH-induced genes yet explain many aspects of Rim101p and PacC function in other organisms.
环境pH值变化对生长和分化具有广泛影响。目前了解最清楚的真核生物pH响应途径是通过构巢曲霉的锌指转录因子PacC起作用,该因子直接激活碱性pH诱导的基因。我们在此表明,酿酒酵母中与PacC同源的pH响应调节因子Rim101p在体内起阻遏物的作用。染色质免疫沉淀试验表明,Rim101p在体内与7个受Rim101p阻遏的基因的启动子相关联。一个含有推导的Rim101p结合位点的报告基因受到Rim101p的负调控,并且在体内与Rim101p相关联。Rim101p阻遏靶标NRG1和SMP1的缺失突变可抑制rim101Δ突变体在离子耐受性、单倍体侵袭性生长和孢子形成方面的缺陷。因此,转录阻遏是Rim101p的主要生物学功能。Rim101p的阻遏靶标Nrg1p反过来又是两个碱性pH诱导基因(包括离子耐受性所需的Na+泵基因ENA1)阻遏所必需的。因此,已知的转录阻遏物Nrg1p作为碱性pH响应的抑制剂发挥作用。我们的发现与PacC作为碱性pH诱导基因的直接激活剂这一已充分表征的功能形成对比,但解释了Rim101p和PacC在其他生物体中的许多功能方面。