Exon junction complex proteins bind nascent transcripts independently of pre-mRNA splicing in Drosophila melanogaster

Elife. 2016 Nov 23:5:e19881. doi: 10.7554/eLife.19881.

Abstract

Although it is currently understood that the exon junction complex (EJC) is recruited on spliced mRNA by a specific interaction between its central protein, eIF4AIII, and splicing factor CWC22, we found that eIF4AIII and the other EJC core proteins Y14 and MAGO bind the nascent transcripts of not only intron-containing but also intronless genes on Drosophila polytene chromosomes. Additionally, Y14 ChIP-seq demonstrates that association with transcribed genes is also splicing-independent in Drosophila S2 cells. The association of the EJC proteins with nascent transcripts does not require CWC22 and that of Y14 and MAGO is independent of eIF4AIII. We also show that eIF4AIII associates with both polysomal and monosomal RNA in S2 cell extracts, whereas Y14 and MAGO fractionate separately. Cumulatively, our data indicate a global role of eIF4AIII in gene expression, which would be independent of Y14 and MAGO, splicing, and of the EJC, as currently understood.

Keywords: D. melanogaster; EJC; chromosomes; genes; splicing; translation.

MeSH terms

  • Animals
  • Cell Fractionation
  • Cells, Cultured
  • Chromosome Mapping
  • Drosophila Proteins / genetics
  • Drosophila Proteins / metabolism
  • Drosophila melanogaster / genetics*
  • Drosophila melanogaster / metabolism
  • Eukaryotic Initiation Factor-4A / genetics*
  • Eukaryotic Initiation Factor-4A / metabolism
  • Exons
  • Introns
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Polytene Chromosomes / chemistry*
  • Polytene Chromosomes / metabolism
  • Protein Binding
  • RNA Precursors / genetics*
  • RNA Precursors / metabolism
  • RNA Splicing*
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism

Substances

  • Drosophila Proteins
  • Mago protein, Drosophila
  • Nuclear Proteins
  • RNA Precursors
  • RNA, Messenger
  • RNA-Binding Proteins
  • tsu protein, Drosophila
  • Eukaryotic Initiation Factor-4A