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Modular function of long noncoding RNA, COLDAIR, in the vernalization response.

Dong-Hwan Kim1, Yanpeng Xi1, Sibum Sung1

  • 1Department of Molecular Biosciences and Institute for Cellular and Molecular Biology, the University of Texas at Austin, TX, United States of America.

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The long noncoding RNA COLDAIR is crucial for plant flowering time control by interacting with PRC2 to repress the FLOWERING LOCUS C gene during cold exposure (vernalization). This interaction ensures proper floral repression.

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Area of Science:

  • Plant Molecular Biology
  • Epigenetics
  • Transcriptomics

Background:

  • Vernalization is a key process regulating flowering time in plants like Arabidopsis thaliana.
  • The floral repressor FLOWERING LOCUS C (FLC) must be repressed for flowering to occur after cold exposure.
  • Polycomb Repressive Complex 2 (PRC2) mediates FLC repression via H3K27me3 epigenetic marks.

Purpose of the Study:

  • To elucidate the molecular mechanism by which the long noncoding RNA COLDAIR interacts with PRC2.
  • To determine the role of COLDAIR in mediating FLC repression during vernalization.
  • To identify the specific regions of COLDAIR critical for PRC2 association and function.

Main Methods:

  • Chromatin immunoprecipitation (ChIP) to assess COLDAIR and PRC2 enrichment at the FLC locus.
  • In vitro binding assays to test COLDAIR-PRC2 interactions.
  • Site-directed mutagenesis of COLDAIR to disrupt the PRC2-binding motif.
  • Analysis of flowering time in Arabidopsis mutants expressing altered COLDAIR.

Main Results:

  • COLDAIR specifically associates with chromatin at the FLC locus.
  • The central region of COLDAIR contains a motif essential for PRC2 binding.
  • Mutations disrupting this motif impair COLDAIR-PRC2 interaction and lead to vernalization insensitivity.
  • Ectopic expression of wild-type COLDAIR rescues the vernalization insensitivity phenotype.

Conclusions:

  • COLDAIR acts as a scaffold, recruiting PRC2 to the FLC locus to mediate epigenetic repression.
  • A specific motif within COLDAIR is critical for its interaction with PRC2.
  • This study reveals the molecular framework of COLDAIR-mediated FLC repression during vernalization.