Global profiling of Rbm24 bound RNAs uncovers a multi-tasking RNA binding protein

Int J Biochem Cell Biol. 2018 Jan:94:10-21. doi: 10.1016/j.biocel.2017.11.002. Epub 2017 Nov 21.

Abstract

RNA binding proteins serve as critical molecular switches in a multitude of post-transcriptional regulatory processes. In the heart and muscles, the tissue specific RNA binding protein, Rbm24, is known to play important developmental roles via driving different post-transcriptional processes. Nonetheless, the currently identified molecular targets and regulatory pathways seem inadequate to completely explain the observed developmental effects upon Rbm24 knockdown/knockout. Here, by performing RNA Immunoprecipitation and coupling it to microarrays (RIP-Chip), we have generated an atlas of the mRNA binding repertoire of Rbm24. Further functional evaluation of its targets led to the elucidation of novel roles for Rbm24 in post-transcriptional processing, besides its already known roles in regulation of mRNA stability and alternative splicing. Interestingly, Rbm24 is found to cause the destabilization of Chrm2 via binding to an element in the coding region. In addition, Rbm24 is also found to have an uncharacterized role in driving the generation of isoforms with alternative transcriptional start sites. We have, for the first time, demonstrated that Rbm24 is a multi-tasking RNA binding protein capable of regulating its bound targets via a range of mechanisms.

Keywords: Alternative splicing; Cardiomyocyte; RIP-Chip; RNA binding protein; Rbm24.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alternative Splicing
  • Animals
  • Binding Sites
  • Cell Line
  • Gene Expression Profiling
  • Genes, Reporter
  • Green Fluorescent Proteins / chemistry
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HEK293 Cells
  • Humans
  • Mice
  • Myocytes, Cardiac / cytology
  • Myocytes, Cardiac / metabolism*
  • Open Reading Frames
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / metabolism
  • Protein Isoforms / chemistry
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • RNA Interference
  • RNA Processing, Post-Transcriptional*
  • RNA Stability
  • RNA, Messenger / chemistry
  • RNA, Messenger / metabolism*
  • RNA-Binding Proteins / antagonists & inhibitors
  • RNA-Binding Proteins / chemistry
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism*
  • Rats
  • Receptor, Muscarinic M2 / chemistry
  • Receptor, Muscarinic M2 / genetics
  • Receptor, Muscarinic M2 / metabolism*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Transcription Initiation Site

Substances

  • CHRM2 protein, human
  • Peptide Fragments
  • Protein Isoforms
  • RBM24 protein, human
  • RNA, Messenger
  • RNA-Binding Proteins
  • Receptor, Muscarinic M2
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Green Fluorescent Proteins