RNA

  • RNA / Volume 7 / Issue 11 / November 2001, pp 1531-1542
  • Copyright © 2001 RNA Society
  • DOI: http://dx.doi.org/ (About DOI), Published online: 11 January 2002


Symmetrical dimethylation of arginine residues in spliceosomal Sm protein B/B′ and the Sm-like protein LSm4, and their interaction with the SMN protein


HERO  BRAHMS a1, LYDIE  MEHEUS a2, VERONIQUE  DE BRABANDERE a2, UTZ  FISCHER a3 and REINHARD  LÜHRMANN a1c1
a1 Max Planck Institute of Biophysical Chemistry, Department of Cellular Biochemistry, 37077 Göttingen, Germany
a2 Innogenetics N.V., Industriepark Zwijnaarde 7, 9052 Ghent, Belgium
a3 Max Planck Institute of Biochemistry, 82152 Martinsried, Germany

Abstract

Arginine residues in RG-rich proteins are frequently dimethylated posttranslationally by protein arginine methyltransferases (PRMTs). The most common methylation pattern is asymmetrical dimethylation, a modification important for protein shuttling and signal transduction. Symmetrically dimethylated arginines (sDMA) have until now been confined to the myelin basic protein MBP and the Sm proteins D1 and D3. We show here by mass spectrometry and protein sequencing that also the human Sm protein B/B′ and, for the first time, one of the Sm-like proteins, LSm4, contain sDMA in vivo. The symmetrical dimethylation of B/B′, LSm4, D1, and D3 decisively influences their binding to the Tudor domain of the “survival of motor neurons” protein (SMN): inhibition of dimethylation by S-adenosylhomocysteine (SAH) abolished the binding of D1, D3, B/B′, and LSm4 to this domain. A synthetic peptide containing nine sDMA-glycine dipeptides, but not asymmetrically modified or nonmodified peptides, specifically inhibited the interaction of D1, D3, B/B′, LSm4, and UsnRNPs with SMN-Tudor. Recombinant D1 and a synthetic peptide could be methylated in vitro by both HeLa cytosolic S100 extract and nuclear extract; however, only the cytosolic extract produced symmetrical dimethylarginines. Thus, the Sm-modifying PRMT is cytoplasmic, and symmetrical dimethylation of B/B′, D1, and D3 is a prerequisite for the SMN-dependent cytoplasmic core-UsnRNP assembly. Our demonstration of sDMAs in LSm4 suggests additional functions of sDMAs in tri-UsnRNP biogenesis and mRNA decay. Our findings also have interesting implications for the understanding of the aetiology of spinal muscular atrophy (SMA).

(Received August 20 2001)
(Revised August 27 2001)
(Accepted September 4 2001)


Key Words: mRNA decay; posttranslational modification; protein arginine methyltransferase (PRMT); protein sequencing; sDMA; spinal muscular atrophy (SMA); spliceosome; UsnRNP.

Correspondence:
c1 Reprint requests to: Reinhard Lührmann, Max Planck Institute of Biophysical Chemistry, Department of Cellular Biochemistry, Am Faßberg 11, 37077 Göttingen, Germany; e-mail: reinhard.luehrmann@mpi-bpc.mpg.de.