Valadi Hadi, Valadi Asa, Ansell Ricky, Gustafsson Lena, Adler Lennart, Norbeck Joakim, Blomberg Anders
Department of Chemistry and Bioscience/Molecular Biotechnology, Chalmers University of Technology, 405-30 Göteborg, Sweden.
Curr Genet. 2004 Feb;45(2):90-5. doi: 10.1007/s00294-003-0469-1. Epub 2003 Dec 2.
A strain of Saccharomyces cerevisiae lacking the GPD2 gene, encoding one of the glycerol-3-phosphate dehydrogenases, grows slowly under anaerobic conditions, due to reductive stress caused by the accumulation of cytoplasmic NADH. We used 2D-PAGE to study the effect on global protein expression of reductive stress in the anaerobically grown gpd2Delta strain. The most striking response was a strongly elevated expression of Tdh1p, the minor isoform of glyceraldehyde-3-phosphate dehydrogenase. This increased expression could be reversed by the addition of acetoin, a NADH-specific redox sink, which furthermore largely restored anaerobic growth of the gpd2Delta strain. Additional deletion of the TDH1 gene (but not of TDH2 or TDH3) improved anaerobic growth of the gpd2Delta strain. We therefore propose that TDH1 has properties not displayed by the other TDH isogenes and that its expression is regulated by reductive stress caused by an excess of cytoplasmic NADH.
一种缺乏编码甘油-3-磷酸脱氢酶之一的GPD2基因的酿酒酵母菌株，在厌氧条件下生长缓慢，这是由于细胞质NADH积累引起的还原应激所致。我们使用二维聚丙烯酰胺凝胶电泳（2D-PAGE）来研究厌氧生长的gpd2Δ菌株中还原应激对整体蛋白质表达的影响。最显著的反应是甘油醛-3-磷酸脱氢酶的次要同工型Tdh1p的表达大幅升高。添加乙偶姻（一种NADH特异性氧化还原阱）可逆转这种增加的表达，而且在很大程度上恢复了gpd2Δ菌株的厌氧生长。额外缺失TDH1基因（而非TDH2或TDH3）改善了gpd2Δ菌株的厌氧生长。因此，我们提出TDH1具有其他TDH同基因未表现出的特性，并且其表达受细胞质NADH过量引起的还原应激调节。
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