Analysis of sDMA Modifications of PIWI Proteins

Methods Mol Biol. 2014;1093:137-48. doi: 10.1007/978-1-62703-694-8_11.

Abstract

Arginine methylation is an important posttranslational protein modification that modulates protein function for a wide range of biological processes. PIWI proteins, a subclade of the Argonaute family proteins, contain evolutionarily conserved symmetrical dimethylarginines (sDMAs). It has become increasingly apparent that the sDMAs of PIWI proteins serve as binding elements for TUDOR domain-containing proteins and that sDMA-dependent protein interactions play crucial roles in the biogenesis and function of PIWI-interacting RNAs (piRNAs). We describe a method for detecting PIWI sDMAs and purifying PIWI/piRNA complexes using anti-sDMA antibodies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Arginine / analogs & derivatives*
  • Arginine / metabolism
  • Argonaute Proteins / chemistry*
  • Argonaute Proteins / isolation & purification
  • Argonaute Proteins / metabolism*
  • Cell Line
  • Electrophoresis, Polyacrylamide Gel
  • Immunoprecipitation
  • Male
  • Mice
  • Protein Processing, Post-Translational*
  • Ribonucleoproteins / metabolism
  • Silver Staining
  • Testis

Substances

  • Argonaute Proteins
  • Ribonucleoproteins
  • dimethylarginine
  • Arginine