Symmetrical dimethylarginine methylation is required for the localization of SMN in Cajal bodies and pre-mRNA splicing

J Cell Biol. 2002 Dec 23;159(6):957-69. doi: 10.1083/jcb.200207028. Epub 2002 Dec 16.

Abstract

The nuclear structures that contain symmetrical dimethylated arginine (sDMA)-modified proteins and the role of this posttranslational modification is unknown. Here we report that the Cajal body is a major epitope in HeLa cells for an sDMA-specific antibody and that coilin is an sDMA-containing protein as analyzed by using the sDMA-specific antibody and matrix-assisted laser desorption ionization time of flight mass spectrometry. The methylation inhibitor 5'-deoxy-5'-methylthioadenosine reduces the levels of coilin methylation and causes the appearance of SMN-positive gems. In cells devoid of Cajal bodies, such as primary fibroblasts, sDMA-containing proteins concentrated in speckles. Cells from a patient with spinal muscular atrophy, containing low levels of the methyl-binding protein SMN, localized sDMA-containing proteins in the nucleoplasm as a discrete granular pattern. Splicing reactions are efficiently inhibited by using the sDMA-specific antibody or by using hypomethylated nuclear extracts, showing that active spliceosomes contain sDMA polypeptides and suggesting that arginine methylation is important for efficient pre-mRNA splicing. Our findings support a model in which arginine methylation is important for the localization of coilin and SMN in Cajal bodies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies / metabolism
  • Arginine / analogs & derivatives
  • Arginine / metabolism*
  • Cell Nucleus / metabolism
  • Coiled Bodies / metabolism*
  • DNA / metabolism
  • Dose-Response Relationship, Drug
  • Down-Regulation
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Fibroblasts / metabolism
  • Fluorescent Antibody Technique, Indirect
  • HeLa Cells
  • Humans
  • Mass Spectrometry
  • Methylation
  • Microscopy, Fluorescence
  • Muscular Atrophy, Spinal / metabolism
  • Precipitin Tests
  • RNA Splicing
  • RNA, Messenger / metabolism
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Transfection

Substances

  • Antibodies
  • RNA, Messenger
  • DNA
  • Arginine