Protein arginine methyltransferase expression and activity during myogenesis

Abstract

Correspondence: Vladimir Ljubicic (ljubicic@mcmaster.ca)Despite the emerging importance of protein arginine methyltransferases (PRMTs) in regulating skeletal muscle plasticity, PRMT biology during muscle development is complex and not completely understood. Therefore, our purpose was to investigate PRMT1, -4, and -5 expression and function in skeletal muscle cells during the phenotypic remodeling elicited by myogenesis. C 2 C 12 muscle cell maturation, assessed during the myoblast (MB) stage, and during days 1, 3, 5, and 7 of differentiation, was employed as an in vitro model of myogenesis. We observed PRMT-specific patterns of expression and activity during myogenesis. PRMT4 and -5 gene expression was unchanged, while PRMT1 mRNA and protein content were significantly induced. Cellular monomethylarginines (MMAs) and symmetric dimethylarginines (SDMAs), indicative of global and type II PRMT activities, respectively, remained steady during development, while type I PRMT activity indicator asymmetric dimethylarginines (ADMAs) increased through myogenesis. Histone 4 arginine 3 (H4R3) and H3R17 contents were elevated coincident with the myonuclear accumulation of PRMT1 and -4. Collectively, this suggests that PRMTs are methyl donors throughout myogenesis and demonstrate specificity for their protein targets. Cells were then treated with TC-E 5003 (TC-E), a selective inhibitor of PRMT1 in order to specifically examine the enzymes role during myogenic differentiation. TC-E treated cells exhibited decrements in muscle differentiation, which were consistent with attenuated mitochondrial biogenesis and respiratory function. In summary, the present study increases our understanding of PRMT1, -4, and -5 biology during the plasticity of skeletal muscle development. Our results provide evidence for a role of PRMT1, via a mitochondrially mediated mechanism, in driving the muscle differentiation program.

Bronchial thermoplasty decreases airway remodelling by blocking epithelium-derived heat shock protein-60 secretion and protein arginine methyltransferase-1 in fibroblasts

“…In this study, exogenous HSP60 increased PRMT1 expression, the corresponding signalling cascade and mitochondrial mass in fibroblasts. Our results indicated that BALF and Epi.S of asthma patients obtained before BT caused mitochondrial dysfunction through upregulation of PGC1α→PRMT1, which is consistent with earlier reports [32,33].…”

Protein arginine methyltransferase biology in humans during acute and chronic skeletal muscle plasticity

“…The following antibodies were used: PRMT1 (07-404, EMD Millipore, Etobicoke, ON, RRID:AB_310588); CARM1 (A300 -421A, Bethyl Laboratories, Montgomery, TX, RRID: AB_420962); PRMT5 (07-405, EMD Millipore, RRID:AB_310589), PRMT7 (sc-376077, Santa Cruz, Dallas, TX, RRID:AB_10990266); PRMT9 (clone 128 -29 -1, EMD Millipore, RRID:AB_2801509); MMA (8015, Cell Signal, Whitby, ON, RRID:AB_2799401); ADMA GAR (13522, Cell Signal, RRID:AB_2665370); CARM1 substrate [denoted as ADMA-5 CARM1 (16), was a kind gift from Dr. Mark Bedford, MD Anderson Cancer Center, University of Texas]; SDMA (13222, Cell Signal, RRID:AB_2714013); histone 4 arginine 3 (H4R3; ab129231, Abcam, Toronto, ON, RRID:AB_2801527); H3R17 (ab8284, Abcam, RRID:AB_306434); H3R8 (ab130740, Abcam, RRID:AB_2801510); H3 (ab1791, Abcam, RRID:AB_302613); and H4 (ab10158, Abcam, RRID:AB_296888). MMA, ADMA GAR , and SDMA are pan-methylation antibodies that have been extensively used in previous literature (16,19,30,48,62,65,71). The ADMA GAR and SDMA antibodies primarily recognize arginine methylation at glycine-and arginine-rich (GAR) motifs.…”

The CARM1 transcriptome and arginine methylproteome mediate skeletal muscle integrative biology

“…CARM1 is also emerging as an important regulator of skeletal muscle biology, a dynamic tissue accounting for 30–40% of body mass that is essential for metabolism, respiration, and mobility [ 12 ]. Indeed, we [ [13] , [14] , [15] , [16] , [17] , [18] , [19] ] and others [ [20] , [21] , [22] , [23] , [24] , [25] ] have shown that CARM1 expression and activity are altered during conditions of muscle plasticity and that the methyltransferase is required for myogenesis, glucose metabolism, and the maintenance and remodelling of muscle mass. In particular, recent work has highlighted CARM1-mediated autophagic and atrophic signalling in skeletal muscle [ 15 , 19 , [24] , [25] , [26] ], revealing a mechanism by which CARM1 influences AMP-activated protein kinase (AMPK) and its downstream network to affect muscle mass.…”