Asymmetric arginine dimethylation of heterogeneous nuclear ribonucleoprotein K by protein-arginine methyltransferase 1 inhibits its interaction with c-Src.
Arginine methylation is a post-translational modification found in many RNA-binding proteins. Heterogeneous nuclear ribonucleoprotein K (hnRNP K) from HeLa cells was shown, by mass spectrometry and Edman degradation, to contain asymmetric N(G),N(G)-dimethylarginine at five positions in its amino acid sequence (Arg256, Arg258, Arg268, Arg296, and Arg299). Whereas these five residues were quantitatively modified, Arg303 was asymmetrically dimethylated in <33% of hnRNP K and Arg287 was monomethylated in <10% of the protein. All other arginine residues were unmethylated. Protein-arginine methyltransferase 1 was identified as the only enzyme methylating hnRNP K in vitro and in vivo. An hnRNP K variant in which the five quantitatively modified arginine residues had been substituted was not methylated. Methylation of arginine residues by protein-arginine methyltransferase 1 did not influence the RNA-binding activity, the translation inhibitory function, or the cellular localization of hnRNP K but reduced the interaction of hnRNP K with the tyrosine kinase c-Src. This led to an inhibition of c-Src activation and hnRNP K phosphorylation. These findings support the role of arginine methylation in the regulation of protein-protein interactions.
Institute of Biochemistry, Martin-Luther-University Halle-Wittenberg, Kurt-Mothes-Strasse 3, 06120 Halle (Saale), Germany. aostareck@biochemtech.uni-halle.de
Antje
A
Ostareck-Lederer
Dirk H
DH
Ostareck
Karl P
KP
Rucknagel
Angelika
A
Schierhorn
Bodo
B
Moritz
Stefan
S
Huttelmaier
Nadine
N
Flach
Lusy
L
Handoko
Elmar
E
Wahle
D000595:Amino Acid Sequence; D001120:Arginine / Q000031:analogs & derivatives / Q000737:chemistry*; D000081247:CSK Tyrosine-Protein Kinase; D019175:DNA Methylation; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian / Q000378:metabolism; D004983:Ethanolamines / Q000737:chemistry; D006367:HeLa Cells; D034641:Heterogeneous-Nuclear Ribonucleoprotein K / Q000737:chemistry*; D006801:Humans; D013058:Mass Spectrometry; D008745:Methylation; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D010766:Phosphorylation; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D014176:Protein Biosynthesis; D025941:Protein Interaction Mapping; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D011484:Protein-Arginine N-Methyltransferases / Q000737:chemistry*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D012313:RNA / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D012097:Repressor Proteins / Q000737:chemistry*; D012685:Sepharose / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013234:Stem Cells / Q000378:metabolism; D014158:Transcription, Genetic; D014162:Transfection; D019061:src-Family Kinases
D004983:Ethanolamines; D034641:Heterogeneous-Nuclear Ribonucleoprotein K; D011994:Recombinant Proteins; D012097:Repressor Proteins; D012313:RNA; C018524:N,N-dimethylarginine; D012685:Sepharose; D001120:Arginine; C099988:PRMT1 protein, human; D011484:Protein-Arginine N-Methyltransferases; D011505:Protein-Tyrosine Kinases; D000081247:CSK Tyrosine-Protein Kinase; D019061:src-Family Kinases; C000631346:CSK protein, human; C007369:2-diethylaminoethanol