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Analysis of sDMA Modifications of PIWI Proteins

  • Shozo Honda
  • Yoriko Kirino
  • Yohei Kirino
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1093)

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.

Key words

PIWI piRNA Arginine methylation Symmetrical dimethylarginine (sDMA) Y12 SYM10 SYM11 

Notes

Acknowledgments

We are grateful to G. Dreyfuss for the Y12 antibody, to S. Katsuma for BmN4 and PIWI expression constructs, and to Z. Mourelatos for support and discussion. This work was supported by the Cedars-Sinai Medical Center Research Fund, Martz Translational Breast Cancer Research Fund, and a Grant for Basic Science Research Project from The Sumitomo Foundation (Y.K.).

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Copyright information

© Springer Science+Business Media, LLC 2014

Authors and Affiliations

  • Shozo Honda
    • 1
  • Yoriko Kirino
    • 1
  • Yohei Kirino
    • 2
  1. 1.Department of Biomedical Sciences, Cedars-Sinai Medical CenterSamuel Oschin Comprehensive Cancer InstituteLos AngelesUSA
  2. 2.Computational Medicine Center, Department of Biochemistry and Molecular BiologyThomas Jefferson UniversityPhiladelphiaUSA

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