Arginine methylation analysis of the splicing-associated SR protein SFRS9/SRP30C.
The human SFRS9/SRp30c belongs to the SR family of splicing regulators. Despite evidence that members of this protein family may be targeted by arginine methylation, this has yet to be experimentally addressed. In this study, we found that SFRS9 is a target for PRMT1-mediated arginine methylation in vitro, and that it is immunoprecipitated from HEK-293 lysates by antibodies that recognize both mono- and dimethylated arginines. We further observed that upon treatment with the methylation inhibitor Adox, the fluorescent EGFP-SFRS9 re-localizes to dot-like structures in the cell nucleus. In subsequent confocal analyses, we found that EGFP-SFRS9 localizes to nucleoli in Adox-treated cells. Our findings indicate the importance of arginine methylation for the subnuclear localization of SFRS9.
Bressan
Gustavo C
GC
Laboratório Nacional de Luz Síncrotron, Campinas, SP, Brasil.
Moraes
Eduardo C
EC
Manfiolli
Adriana O
AO
Kuniyoshi
Tais M
TM
Passos
Dario O
DO
Gomes
Marcelo D
MD
Kobarg
Jörg
J
D021581:Active Transport, Cell Nucleus; D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology; D000595:Amino Acid Sequence; D001120:Arginine / Q000032:analysis* / Q000378:metabolism; D002460:Cell Line; D006801:Humans; D008745:Methylation / Q000187:drug effects; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012326:RNA Splicing; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D000068103:Serine-Arginine Splicing Factors
D009687:Nuclear Proteins; D020033:Protein Isoforms; D016601:RNA-Binding Proteins; D000068103:Serine-Arginine Splicing Factors; C027579:periodate-oxidized adenosine; D001120:Arginine; D000241:Adenosine