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CARM1 methylates MED12 to regulate its RNA-binding ability.

Donghang Cheng1, Vidyasiri Vemulapalli1, Yue Lu1

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Coactivator-associated arginine methyltransferase (CARM1) methylates MED12, a key protein in enhancer function. This methylation is crucial for estrogen-regulated gene transcription and interactions with noncoding RNAs.

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Area of Science:

  • Epigenetics and Gene Regulation
  • Molecular Biology
  • Biochemistry

Background:

  • Coactivator-associated arginine methyltransferase (CARM1) is a key transcriptional regulator.
  • CARM1's diverse substrate methylation impacts cellular processes.
  • Understanding CARM1's full substrate repertoire is essential for elucidating its regulatory mechanisms.

Purpose of the Study:

  • To identify novel CARM1 substrates and elucidate their roles in gene regulation.
  • To investigate the specific methylation site on MED12 and its functional consequences.
  • To explore the interaction between CARM1, MED12, and noncoding RNAs in transcriptional control.

Main Methods:

  • Generation of CARM1 substrate motif antibodies.
  • Immunoprecipitation coupled with mass spectrometry to identify CARM1 targets.
  • Chromatin immunoprecipitation sequencing (ChIP-seq) to analyze CARM1 localization and function.
  • Site-directed mutagenesis to investigate the MED12 R1899 methylation site.

Main Results:

  • Identification of MED12 and KMT2D as novel CARM1 substrates.
  • Pinpointing arginine 1899 (R1899) on MED12 as the primary CARM1 methylation site.
  • Demonstration that CARM1 and its MED12 methylation mark are enriched at ERα-specific enhancers.
  • Evidence that MED12 methylation at R1899 and TDRD3 recruitment are vital for MED12's interaction with activating noncoding RNAs.
  • CARM1 positively modulates transcription of estrogen-regulated genes.

Conclusions:

  • CARM1-mediated methylation of MED12 at R1899 is a critical regulatory event in transcription.
  • This methylation event influences enhancer function and estrogen-regulated gene expression.
  • The MED12-TDRD3 interaction, mediated by CARM1, is essential for noncoding RNA-mediated transcriptional activation.