Rai Rajendra, Tate Jennifer J, Cooper Terrance G
Department of Molecular Sciences, University of Tennessee, Memphis, Tennessee 38163, USA.
J Biol Chem. 2003 Apr 11;278(15):12826-33. doi: 10.1074/jbc.M212186200. Epub 2003 Jan 31.
Ure2, the protein that negatively regulates GATA factor (Gln3, Gat1)-mediated transcription in Saccharomyces cerevisiae, possesses prion-like characteristics. Identification of metabolic and environmental factors that influence prion formation as well as any activities that prions or prion precursors may possess are important to understanding them and developing treatment strategies for the diseases in which they participate. Ure2 exhibits primary sequence and three-dimensional homologies to known glutathione S-transferases. However, multiple attempts over nearly 2 decades to demonstrate Ure2-mediated S-transferase activity have been unsuccessful, leading to the possibility that Ure2 may well not participate in glutathionation reactions. Here we show that Ure2 is required for detoxification of glutathione S-transferase substrates and cellular oxidants. ure2 Delta mutants are hypersensitive to cadmium and nickel ions and hydrogen peroxide. They are only slightly hypersensitive to diamide, which is nitrogen source-dependent, and minimally if at all hypersensitive to 1-chloro-2,4-dinitrobenzene, the most commonly used substrate for glutathione S-transferase enzyme assays. Therefore, Ure2 shares not only structural homology with various glutathione S-transferases, but ure2 mutations possess the same phenotypes as mutations in known S. cerevisiae and Schizosaccharomyces pombe glutathione S-transferase genes. These findings are consistent with Ure2 serving as a glutathione S-transferase in S. cerevisiae.
Ure2蛋白在酿酒酵母中负调控GATA因子(Gln3、Gat1)介导的转录,具有朊病毒样特征。确定影响朊病毒形成的代谢和环境因素以及朊病毒或朊病毒前体可能具有的任何活性,对于了解它们以及制定针对它们所参与疾病的治疗策略至关重要。Ure2与已知的谷胱甘肽S-转移酶具有一级序列和三维同源性。然而,近20年来多次尝试证明Ure2介导的S-转移酶活性均未成功,这使得Ure2很可能不参与谷胱甘肽化反应。在此我们表明,Ure2是谷胱甘肽S-转移酶底物和细胞氧化剂解毒所必需的。ure2Δ突变体对镉、镍离子和过氧化氢高度敏感。它们对二酰胺仅略有超敏反应,二酰胺的超敏反应依赖于氮源,而对谷胱甘肽S-转移酶酶活性测定中最常用的底物1-氯-2,4-二硝基苯几乎没有超敏反应(如果有的话)。因此,Ure2不仅与各种谷胱甘肽S-转移酶具有结构同源性,而且ure2突变与已知的酿酒酵母和粟酒裂殖酵母谷胱甘肽S-转移酶基因突变具有相同的表型。这些发现与Ure2在酿酒酵母中作为谷胱甘肽S-转移酶的作用一致。