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Crosstalk between the RNA Methylation and Histone-Binding Activities of MePCE Regulates P-TEFb Activation on

Samantha B Shelton1, Nakul M Shah1, Nathan S Abell2

  • 1Department of Molecular Biosciences, 2500 Speedway, Austin, TX 78712, USA.

Cell Reports
|February 10, 2018
PubMed

Insights

MePCE protein activates transcription by binding chromatin, not just RNA. This interaction releases P-TEFb kinase, enabling gene expression crucial for cell identity.

Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Epigenetics

Background:

  • RNA polymerase II (RNAP II) transcription elongation is regulated by P-TEFb kinase.
  • P-TEFb activity is inhibited by the 7SK small nuclear ribonucleoprotein (7SK snRNP) complex in the nucleosol.
  • MePCE is an RNA methyltransferase essential for 7SK snRNP stability.

Purpose of the Study:

  • To investigate the role of MePCE in regulating P-TEFb activity and transcription elongation.
  • To elucidate the mechanism by which MePCE controls P-TEFb localization and function.
  • To understand how MePCE's dual activities (RNA methylation and chromatin binding) impact gene regulation.

Main Methods:

  • Chromatin immunoprecipitation (ChIP) assays to detect MePCE binding to chromatin.
  • In vitro assays to assess MePCE's RNA methyltransferase activity and histone binding.
  • Biochemical assays to measure P-TEFb kinase activity in different cellular contexts.

Main Results:

  • MePCE binds to chromatin via histone H4 tails, independent of 7SK snRNP.
  • Chromatin-bound MePCE acts as a P-TEFb activator at specific genes.
  • Histone binding inhibits MePCE's RNA methyltransferase activity, releasing P-TEFb for RNAP II activation.
  • MePCE-mediated P-TEFb activation on chromatin is independent of 7SK snRNP and Brd4.

Conclusions:

  • MePCE's dual functions in RNA methylation and chromatin binding are critical for regulating P-TEFb activity.
  • MePCE acts as a molecular switch, modulating P-TEFb availability for transcription based on its interaction with chromatin.
  • This mechanism provides a novel pathway for regulating gene expression important for cellular identity, independent of the canonical 7SK snRNP pathway.
Keywords:
7SKBCDIN3MePCERNA methyltransferaseRNAP II pausingchromatinhistonesnon-coding RNApTEFb

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