Nuclear accumulation of mRNAs underlies G4C2-repeat-induced translational repression in a cellular model of C9orf72 ALS

J Cell Sci. 2015 May 1;128(9):1787-99. doi: 10.1242/jcs.165332. Epub 2015 Mar 18.

Abstract

A common feature of non-coding repeat expansion disorders is the accumulation of RNA repeats as RNA foci in the nucleus and/or cytoplasm of affected cells. These RNA foci can be toxic because they sequester RNA-binding proteins, thus affecting various steps of post-transcriptional gene regulation. However, the precise step that is affected by C9orf72 GGGGCC (G4C2) repeat expansion, the major genetic cause of amyotrophic lateral sclerosis (ALS), is still poorly defined. In this work, we set out to characterise these mechanisms by identifying proteins that bind to C9orf72 RNA. Sequestration of some of these factors into RNA foci was observed when a (G4C2)31 repeat was expressed in NSC34 and HeLa cells. Most notably, (G4C2)31 repeats widely affected the distribution of Pur-alpha and its binding partner fragile X mental retardation protein 1 (FMRP, also known as FMR1), which accumulate in intra-cytosolic granules that are positive for stress granules markers. Accordingly, translational repression is induced. Interestingly, this effect is associated with a marked accumulation of poly(A) mRNAs in cell nuclei. Thus, defective trafficking of mRNA, as a consequence of impaired nuclear mRNA export, might affect translation efficiency and contribute to the pathogenesis of C9orf72 ALS.

Keywords: Amyotrophic lateral sclerosis; C9orf72; Stress granules; mRNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amyotrophic Lateral Sclerosis / metabolism*
  • Amyotrophic Lateral Sclerosis / pathology
  • Animals
  • C9orf72 Protein
  • Cell Nucleus / metabolism*
  • DNA-Binding Proteins
  • Eukaryotic Initiation Factor-2 / metabolism
  • Fragile X Mental Retardation Protein / metabolism
  • HeLa Cells
  • Humans
  • Intracellular Space / metabolism
  • Mice
  • Models, Biological*
  • Motor Neurons / metabolism
  • Phosphorylation
  • Poly(A)-Binding Proteins / metabolism
  • Protein Binding
  • Protein Biosynthesis*
  • Proteins / metabolism*
  • RNA Splicing / genetics
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Transcription Factors
  • Trinucleotide Repeat Expansion*

Substances

  • C9orf72 Protein
  • C9orf72 protein, human
  • DNA-Binding Proteins
  • Eukaryotic Initiation Factor-2
  • PURA protein, human
  • Poly(A)-Binding Proteins
  • Proteins
  • RNA, Messenger
  • Transcription Factors
  • Fragile X Mental Retardation Protein