Arginine methylation in yeast proteins during stationary-phase growth and heat shock.

Abstract

Arginine methyltransferases (RMTs) catalyze the methylation of arginine residues on proteins. We examined the effects of log-phase growth, stationary-phase growth, and heat shock on the formation of methylarginines on yeast proteins to determine if the conditions favor a particular type of methylation. Utilizing linear ion trap mass spectrometry, we identify methylarginines in wild-type and RMT deletion yeast strains using secondary product ion scans (MS(3)), and quantify the methylarginines using multiple reaction monitoring (MRM). Employing MS(3) and isotopic incorporation, we demonstrate for the first time that Nη1, Nη2-dimethylarginine (sDMA) is present on yeast proteins, and make a detailed structural determination of the fragment ions from the spectra. Nη-monomethylarginine (ηMMA), Nδ-monomethylarginine (δMMA), Nη1, Nη1-dimethylarginine (aDMA), and sDMA were detected in RMT deletion yeast using MS(3) and MRM with and without isotopic incorporation, suggesting that additional RMT enzymes remain to be discovered in yeast. The concentrations of ηMMA and δMMA decreased by half during heat shock and stationary phase compared to log-phase growth of wild-type yeast, whereas sDMA increased by as much as sevenfold and aDMA decreased by 11-fold. Therefore, upon entering stressful conditions like heat shock or stationary-phase growth, there is a net increase in sDMA and decreases in aDMA, ηMMA, and δMMA on yeast proteins.

• Authors: Ted M Lakowski, Magnolia L Pak, András Szeitz, Dylan Thomas, Mynol I Vhuiyan, Bernd Clement, Adam Frankel
• Journal: Amino acids (Amino Acids) Vol. 47 Issue 12 Pg. 2561-71 (Dec 2015) ISSN: 1438-2199 [Electronic] Austria
• PMID: 26189025 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Saccharomyces cerevisiae Proteins
• dimethylarginine
• omega-N-Methylarginine
• Arginine
• Protein-Arginine N-Methyltransferases
• HSL7 protein, S cerevisiae

Topics

• Arginine (analogs & derivatives, chemistry)
• Gene Deletion
• Heat-Shock Response
• Hot Temperature
• Mass Spectrometry
• Methylation
• Protein-Arginine N-Methyltransferases (chemistry)
• Saccharomyces cerevisiae (chemistry)
• Saccharomyces cerevisiae Proteins (chemistry)
• omega-N-Methylarginine (chemistry)