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REST interacts with Cbx proteins and regulates polycomb repressive complex 1 occupancy at RE1 elements.

Xiaojun Ren1, Tom K Kerppola

  • 1Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor, MI 48109-0650, USA.

Molecular and Cellular Biology
|March 16, 2011
PubMed
Summary

REST and Polycomb group (PcG) proteins interact to regulate gene expression during cell differentiation. Their opposing effects on PcG binding at different DNA elements enable precise control of neuronal gene transcription.

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Area of Science:

  • Epigenetics
  • Developmental Biology
  • Molecular Biology

Background:

  • Polycomb group (PcG) proteins are crucial for epigenetic inheritance of transcriptional states during development.
  • Cellular differentiation involves coordinated activation and repression of PcG target genes.

Purpose of the Study:

  • To investigate the interaction between REST and PcG proteins in mouse embryonic stem cells.
  • To elucidate the role of REST in modulating PcG complex binding and function at regulatory elements.

Main Methods:

  • Co-immunoprecipitation assays to identify interacting proteins.
  • In vivo and in vitro studies of REST-Cbx and REST-Ring1b interactions.
  • Chromatin immunoprecipitation to assess PcG binding at REST-bound elements.
  • Gene expression analysis following REST or PcG subunit manipulation.

Main Results:

  • REST and associated proteins were found to copurify with Cbx family PcG proteins in mouse ES cells.
  • REST interacted with Cbx proteins and endogenous Ring1b, with PRC1 subunits occupying REST-bound sites.
  • Manipulation of REST levels altered PRC1 binding at proximal and distal RE1 elements, leading to differential gene transcription effects.
  • Similar effects were observed for REST and PRC1 subunit knockdowns/knockouts on neuronal gene expression.

Conclusions:

  • REST influences PcG occupancy at distinct regulatory elements, exhibiting opposite effects on proximal versus distal sites.
  • These context-dependent actions of REST on PcG binding contribute to the gene-specific regulation of PcG functions during ES cell differentiation.