Ribosomal protein S2 is a substrate for mammalian PRMT3 (protein arginine methyltransferase 3).
PRMT3 (protein arginine methyltransferase 3) is one of four type I arginine methyltransferases that catalyse the formation of asymmetric dimethylarginine. PRMT3 is unique in that its N-terminus harbours a C2H2 zinc-finger domain that is proposed to confer substrate specificity. In addition, PRMT3 is the only type I enzyme that is restricted to the cytoplasm. Known in vitro substrates for PRMT3 include GST-GAR (a glutathione S-transferase fusion protein containing the glycine- and arginine-rich N-terminal region of fibrillarin), Sam68 (Src-associated substrate during mitosis 68 kDa) and PABP-N1 [poly(A)-binding protein-N1; PABP2]. Here we report the identification of an in vivo substrate for mammalian PRMT3. We found that FLAG-tagged PRMT3 can 'pull down' a protein with a molecular mass of 30 kDa from HeLa cell extracts. MS identified this PRMT3-interacting protein as rpS2 (ribosomal protein S2). In vitro studies showed that the zinc-finger domain of PRMT3 is necessary and sufficient for binding to rpS2. In addition, rpS2 is methylated by PRMT3 in vitro and is also methylated in cell lines. Deletion analysis of the rpS2 amino acid sequence identified a N-terminal Arg-Gly repeat as the methylation site. Furthermore, both PRMT3 and rpS2 co-sediment with free ribosomal subunits. These studies implicate PRMT3 in ribosomal function and in the regulation of protein synthesis.
The University of Texas M.D. Anderson Cancer Center, Science Park-Research Division, P.O. Box 389, Smithville, TX 78957, USA.
Rafal
R
Swiercz
Maria D
MD
Person
Mark T
MT
Bedford
D000818:Animals; D001120:Arginine / Q000378:metabolism; D002846:Chromatography, Affinity; D006367:HeLa Cells; D006801:Humans; D008745:Methylation; D051379:Mice; D041681:NIH 3T3 Cells; D014176:Protein Biosynthesis / Q000502:physiology*; D025941:Protein Interaction Mapping; D011484:Protein-Arginine N-Methyltransferases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012269:Ribosomal Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism; D016335:Zinc Fingers / Q000502:physiology
D011993:Recombinant Fusion Proteins; D012269:Ribosomal Proteins; C030004:ribosomal protein S2; D001120:Arginine; C502438:PRMT3 protein, mouse; D011484:Protein-Arginine N-Methyltransferases