PRMT3 inhibits ubiquitination of ribosomal protein S2 and together forms an active enzyme complex.
Protein arginine methyltransferase 3 (PRMT3) comprises a region not required for catalytic activity in its amino-terminus and the core domain catalyzing protein arginine methylation. PRMT3 has been shown to interact with the 40S ribosomal protein S2 (rpS2) and methylate arginine residues in the arginine-glycine (RG) repeat region in the amino-terminus of rpS2. We investigated the biological implications of this interaction by delineating the domains that mediate binding between PRMT3 and rpS2. The rpS2 (100-293 amino acids) domain, but not the amino-terminus of rpS2 that includes the RG repeat region was essential for binding to PRMT3 and was susceptible to degradation. The amino-terminus of PRMT3, but not its catalytic core was required for binding to and the stability of rpS2. Overexpressed rpS2 was ubiquitinated in cells, but expression of PRMT3 reduced this ubiquitination and stabilized the rpS2 protein. Recombinant PRMT3 formed an active enzyme complex with endogenous rpS2 in vitro. Recombinant rpS2 in molar excess modestly increased the enzymatic activity of PRMT3 in vitro. Our results suggest that in addition to its catalytic function, PRMT3 may control the level of rpS2 protein in cells by inhibiting ubiquitin-mediated proteolysis of rpS2, while rpS2 may regulate the enzymatic activity of PRMT3 as a likely non-catalytic subunit.
Choi
Seeyoung
S
Gene Therapy Research Unit, KRIBB, Yuseong, Daejeon 305-806, Republic of Korea.
Jung
Cho-Rok
CR
Kim
Jin-Young
JY
Im
Dong-Soo
DS
D002384:Catalysis; D020134:Catalytic Domain; D002460:Cell Line; D006801:Humans; D009097:Multienzyme Complexes / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D011484:Protein-Arginine N-Methyltransferases / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012269:Ribosomal Proteins / Q000737:chemistry / Q000378:metabolism*; D013816:Thermodynamics; D054875:Ubiquitination
D009097:Multienzyme Complexes; D011993:Recombinant Fusion Proteins; D012269:Ribosomal Proteins; C030004:ribosomal protein S2; D011484:Protein-Arginine N-Methyltransferases