title: 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.affiliation: Pharmaceutical Analysis Laboratory, Faculty of Health Sciences, College of Pharmacy, The University of Manitoba, 750 McDermot Avenue, Winnipeg, Manitoba, R3E 0T5, Canada. Ted.Lakowski@umanitoba.ca.
authors.forename: Ted M
authors.initials: TM
authors.lastname: Lakowski
authors.affiliation: University of Massachusetts Medical School, Worcester, MA, USA.
authors.forename: Magnolia L
authors.initials: ML
authors.lastname: Pak
authors.affiliation: Faculty of Pharmaceutical Sciences, The University of British Columbia, 2405 Wesbrook Mall, Vancouver, BC, V6T 1Z3, Canada.
authors.forename: András
authors.initials: A
authors.lastname: Szeitz
authors.affiliation: Faculty of Pharmaceutical Sciences, The University of British Columbia, 2405 Wesbrook Mall, Vancouver, BC, V6T 1Z3, Canada.
authors.forename: Dylan
authors.initials: D
authors.lastname: Thomas
authors.affiliation: Faculty of Pharmaceutical Sciences, The University of British Columbia, 2405 Wesbrook Mall, Vancouver, BC, V6T 1Z3, Canada.
authors.forename: Mynol I
authors.initials: MI
authors.lastname: Vhuiyan
authors.affiliation: Pharmaceutical Institute, Gutenbergstr. 76, 24118, Kiel, Germany.
authors.forename: Bernd
authors.initials: B
authors.lastname: Clement
authors.affiliation: Faculty of Pharmaceutical Sciences, The University of British Columbia, 2405 Wesbrook Mall, Vancouver, BC, V6T 1Z3, Canada. afrankel@mail.ubc.ca.
authors.forename: Adam
authors.initials: A
authors.lastname: Frankel
mesh_terms: D001120:Arginine / Q000031:analogs & derivatives / Q000737:chemistry*; D017353:Gene Deletion; D018869:Heat-Shock Response; D006358:Hot Temperature; D013058:Mass Spectrometry; D008745:Methylation; D011484:Protein-Arginine N-Methyltransferases / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000737:chemistry*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry*; D019323:omega-N-Methylarginine / Q000737:chemistry
chemical_list: D029701:Saccharomyces cerevisiae Proteins; C487735:dimethylarginine; D019323:omega-N-Methylarginine; D001120:Arginine; D011484:Protein-Arginine N-Methyltransferases; C120666:HSL7 protein, S cerevisiae
keywords: ADMA; Delta-monomethylarginine; Heat shock; Log phase; MRM3; Mass spectrometry; Protein methylation; Stationary phase; Yeast; sDMA