A Homologue of Elongation Factor 1γ Regulates Methionine Sulfoxide Reductase A Gene Expression in Saccharomyces cerevisiae

• Published in: Proceedings of the National Academy of Sciences - PNAS 2003-07, Vol.100 (14), p.8199-8204
• Main Authors: Hanbauer, Ingeborg, Boja, Emily S., Moskovitz, Jackob
• Format: Article
• Language: English
• Subjects: Antibodies; Biological Sciences; DNA; DNA, Fungal - genetics; Gene expression; Gene Expression Regulation, Fungal; Genes; Methionine - analogs & derivatives; Methionine - metabolism; Methionine Sulfoxide Reductases; Nuclear proteins; Oxidation-Reduction; Oxidative Stress; Oxidoreductases - biosynthesis; Oxidoreductases - genetics; Peptide Elongation Factor 1 - genetics; Peptide Elongation Factors - deficiency; Peptide Elongation Factors - genetics; Peptide Elongation Factors - physiology; Promoter Regions, Genetic; Protein Binding; Protein Structure, Tertiary; Quaternary ammonium compounds; RNA; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - physiology; Sulfates; Sulfoxides; TATA Box; Transcription, Genetic; Yeasts
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.1432898100

Methionine sulfoxide reductase A (MsrA) maintains the function of many proteins by reversing oxidation of methionine residues. Lack of this repair mechanism very likely increases aging-related disease susceptibility. In Saccharomyces cerevisiae, disruption of the msrA gene increases free and protein-bound methionine sulfoxide and decreases cell viability. Although the underlying mechanisms in the induction of the msrA gene are still unknown, a transcriptional regulation may be involved. Hence, a search of nuclear proteins regulating the msrA gene is a major target of the experiments reported in this article. Using protein purification combined with MS, we discovered that calcium phospholipid-binding protein (CPBP), a homologue of elongation factor-1γ, is a component of a complex that binds to the msrA promoter. By measuring CPBP cooperative binding to the msrA promoter, we have mapped the CPBP binding site to a 39-bp sequence at the 3′ end of the promoter. In a mutant yeast strain lacking the CPBP-encoding gene, the ability to overexpress msrA mRNA and MsrA protein was impaired and MsrA catalytic activity was greatly reduced, suggesting that CPBP may enhance msrA gene expression.