Methylglyoxal is a ubiquitous 2-oxoaldehyde derived from glycolysis. Although an endogenous metabolite, methylglyoxal at high concentrations has deleterious effects on cellular functions. Since pretreatment of Saccharomyces cerevisiae cells with methylglyoxal at a low concentration alleviates the toxicity of a subsequent lethal concentration of this 2-oxoaldehyde, proteins synthesized during treatment with methylglyoxal are necessary for adaptation to methylglyoxal. Nevertheless, here we show that methylglyoxal attenuates the rate of overall protein synthesis in S. cerevisiae. Phosphorylation of the alpha subunit of translation initiation factor 2 (eIF2 alpha) is induced by several types of environmental stress, and subsequently, overall protein synthesis is reduced due to the impairment of the formation of a translation initiation complex. We found that methylglyoxal activates the protein kinase Gcn2 to phosphorylate eIF2 alpha. The transcription factor Gcn4 is a master regulator of gene expression under conditions of amino acid starvation and some environmental stresses, the level of which is regulated by Gcn2. We found that adaptation to methylglyoxal was impaired in gcn4 Delta cells, indicating the expression of certain genes regulated by Gcn4 to be important for the adaptive response to methylglyoxal. (C) 2008 Elsevier Inc. All rights reserved.