Asymmetric Arginine Dimethylation of Heterogeneous Nuclear Ribonucleoprotein K by Protein-arginine Methyltransferase 1 Inhibits Its Interaction with c-Src*

  1. Antje Ostareck-Lederer ‡ 1 ,
  2. Dirk H. Ostareck ‡ ,
  3. Karl P. Rucknagel § ,
  4. Angelika Schierhorn ‡ ,
  5. Bodo Moritz ‡ ,
  6. Stefan Huttelmaier ,
  7. Nadine Flach ‡ ,
  8. Lusy Handoko ∥ and
  9. Elmar Wahle ‡ 2
  1. ‡Institute of Biochemistry, Martin-Luther-University Halle-Wittenberg, Kurt-Mothes-Strasse 3, 06120 Halle (Saale), the §Max Planck Research Unit for Enzymology of Protein Folding, Weinbergweg 22, 06120 Halle (Saale), ZAMED, Martin-Luther-University Halle-Wittenberg, Heinrich Damerow Strasse 1, 06120 Halle (Saale), and the ∥Institute of Biochemistry, Biocenter at the University of Wuerzburg, Am Hubland, 97074 Wuerzburg, Germany
  1. 1 To whom correspondence should be addressed. Tel.: 49-(0)345-552-4949; Fax: 49-(0)345-552-7014; E-mail: aostareck{at}biochemtech.uni-halle.de.

Abstract

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 NG,NG-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.

Footnotes

  • ↵3 The abbreviations used are: PRMT, protein-arginine methyltransferase; hnRNP K, heterogeneous ribonucleoprotein K; ES, embryonic stem; Hrp1, nuclear polyadenylated RNA-binding protein 4 (Nap4p); Sam68, Src associated in mitosis 68-kDa protein; PABPN1, mammalian nuclear poly(A)-binding protein 1; r15-LOX, reticulocyte-15-lipoxygenase; DICE, differentiation control element; L2, protein L2 of human papillomavirus type 16; GST, glutathione S-transferase; LIF, leukemia inhibitory factor; GST-GAR, GST fusion protein containing the glycine- and arginine-rich N-terminal region of fibrillarin; HPLC, high pressure liquid chromatography; MALDI-TOF MS, matrix-assisted laser desorption ionization-time of flight mass spectrometry; ESI MS, electrospray ionization-mass spectrometry; SAM, S-adenosylmethionine; SH3-domain, Src homology-3 domain; Src, tyrosine kinase of the Rous sarcoma virus; WW, denotes two conserved tryptophan residues within this domain; Erk, extracellular signal-regulated kinase; GFP, green fluorescent protein; MS/MS, tandem mass spectrometry.

  • ↵4 A. Ostareck-Lederer and D. H. Ostareck, unpublished results.

  • ↵* This work was supported in part by a Heisenberg fellowship of the Deutsche Forschungsgemeinschaft (DFG) (to A. O.-L.) and by the DFG (Grants Os 290/2-1 to A. O.-L., Os 135/2-1,2 to D. H. O. and Grant SFB 610 to E. W.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

  • ↵Graphic The on-line version of this article (available at http://www.jbc.org) contains supplemental text, Ref. 1, and Fig. S1.

  • ↵2 Supported by the Fonds der Chemischen Industrie.

    • Received December 7, 2005.
    • Revision received February 2, 2006.
View this article with LENS
Table of Contents

This Article

  1. First Published on February 21, 2006 doi: 10.1074/jbc.M513053200 The Journal of Biological Chemistry 281, 11115-11125.
  1. » Abstract(Free)
  2. Full Text(Free)
  3. PDF(Free)
  4. PDF including Supp Data
  5. Supplemental Data
  6. All Versions of this Article:
    1. M513053200v1
    2. 281/16/11115 (most recent)

Article Usage Stats

  1. Article Usage Statistics

Submit your work to JBC.

You'll be in good company.