Automethylation of CARM1 allows coupling of transcription and mRNA splicing.
Coactivator-associated arginine methyltransferase 1 (CARM1), the histone arginine methyltransferase and coactivator for many transcription factors, is subject to multiple post-translational modifications (PTMs). To unbiasedly investigate novel CARM1 PTMs we employed high-resolution top-down mass spectrometry. Surprisingly, mouse CARM1 expressed in insect and mammalian expression systems was completely dimethylated at a single site in the C-terminal domain (CTD). We demonstrate that dimethylation of CARM1 occurs both in vivo and in vitro and proceeds via an automethylation mechanism. To probe function of automethylation, we mutated arginine 551 to lysine to create an automethylation-deficient CARM1. Although mutation of CARM1's automethylation site did not affect its enzymatic activity, it did impair both CARM1-activated transcription and pre-mRNA splicing. These results strongly imply that automethylation of CARM1 provides a direct link to couple transcription and pre-mRNA splicing in a manner differing from the other steroid receptor coactivators. Furthermore, our study identifies a self-regulatory signaling mechanism from CARM1's catalytic domain to its CTD.
McArdle Laboratory for Cancer Research and Human Proteomics Program, School of Medicine and Public Health, University of Wisconsin, Madison, WI 53706, USA.
Peter
P
Kuhn
Rob
R
Chumanov
Yidan
Y
Wang
Ying
Y
Ge
Richard R
RR
Burgess
Wei
W
Xu
D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000737:chemistry; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006801:Humans; D008745:Methylation; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D011484:Protein-Arginine N-Methyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012326:RNA Splicing; D012333:RNA, Messenger / Q000378:metabolism*; D014158:Transcription, Genetic
D012333:RNA, Messenger; D001120:Arginine; D011484:Protein-Arginine N-Methyltransferases; C119966:coactivator-associated arginine methyltransferase 1