ATP-dependent roX RNA remodeling by the helicase maleless enables specific association of MSL proteins

Mol Cell. 2013 Jul 25;51(2):174-84. doi: 10.1016/j.molcel.2013.06.011.

Abstract

Dosage compensation in Drosophila involves a global activation of genes on the male X chromosome. The activating complex (MSL-DCC) consists of male-specific-lethal (MSL) proteins and two long, noncoding roX RNAs. The roX RNAs are essential for X-chromosomal targeting, but their contributions to MSL-DCC structure and function are enigmatic. Conceivably, the RNA helicase MLE, itself an MSL subunit, is actively involved in incorporating roX into functional DCC. We determined the secondary structure of roX2 and mapped specific interaction sites for MLE in vitro. Upon addition of ATP, MLE disrupted a functionally important stem loop in roX2. This RNA remodeling enhanced specific ATP-dependent association of MSL2, the core subunit of the MSL-DCC, providing a link between roX and MSL subunits. Probing the conformation of roX in vivo revealed a remodeled stem loop in chromatin-bound roX2. The active remodeling of a stable secondary structure by MLE may constitute a rate-limiting step for MSL-DCC assembly.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / pharmacology*
  • Animals
  • Animals, Genetically Modified
  • Base Pairing
  • Blotting, Western
  • Chromatin / genetics
  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / metabolism*
  • DNA Helicases / genetics
  • DNA Helicases / metabolism*
  • Drosophila Proteins / chemistry
  • Drosophila Proteins / genetics*
  • Drosophila Proteins / metabolism*
  • Drosophila melanogaster / genetics
  • Drosophila melanogaster / growth & development
  • Drosophila melanogaster / metabolism
  • Electrophoretic Mobility Shift Assay
  • Genes, DCC / genetics
  • Immunoprecipitation
  • Male
  • Mutation / genetics
  • Nucleic Acid Conformation
  • RNA / chemistry
  • RNA / genetics*
  • RNA / metabolism
  • RNA Helicases / genetics
  • RNA Helicases / metabolism*
  • RNA-Binding Proteins / chemistry
  • RNA-Binding Proteins / genetics*
  • RNA-Binding Proteins / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Transcription Factors / chemistry
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism*
  • Transcription, Genetic
  • X Chromosome / genetics*
  • X Chromosome / metabolism

Substances

  • Chromatin
  • Chromosomal Proteins, Non-Histone
  • Drosophila Proteins
  • Pabp2 protein, Drosophila
  • RNA-Binding Proteins
  • Recombinant Proteins
  • Transcription Factors
  • mle protein, Drosophila
  • roX1 protein, Drosophila
  • RNA
  • Adenosine Triphosphate
  • DNA Helicases
  • RNA Helicases