Ribosome profiling reveals novel regulation of C9ORF72 GGGGCC repeat-containing RNA translation

RNA. 2022 Feb;28(2):123-138. doi: 10.1261/rna.078963.121. Epub 2021 Nov 30.

Abstract

GGGGCC (G4C2) repeat expansion in the first intron of C9ORF72 causes amyotrophic lateral sclerosis and frontotemporal dementia. Repeat-containing RNA is translated into dipeptide repeat (DPR) proteins, some of which are neurotoxic. Using dynamic ribosome profiling, we identified three translation initiation sites in the intron upstream of (G4C 2) repeats; these sites are detected irrespective of the presence or absence of the repeats. During translocation, ribosomes appear to be stalled on the repeats. An AUG in the preceding C9ORF72 exon initiates a uORF that inhibits downstream translation. Polysome isolation indicates that unspliced (G4C 2) repeat-containing RNA is a substrate for DPR protein synthesis. (G4C 2) repeat-containing RNA translation is 5' cap-independent but inhibited by the initiation factor DAP5, suggesting an interplay with uORF function. These results define novel translational mechanisms of expanded (G4C 2) repeat-containing RNA in disease.

Keywords: ALS; C9ORF72; DAP5; frontotemporal dementia; ribosome profiling; ribosome stalling; uORF translation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • C9orf72 Protein / genetics*
  • C9orf72 Protein / metabolism
  • Dinucleotide Repeats
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / metabolism
  • Neural Stem Cells / cytology
  • Neural Stem Cells / metabolism
  • Peptide Chain Initiation, Translational*
  • RNA, Messenger / chemistry*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Ribosomes / metabolism*

Substances

  • C9orf72 Protein
  • C9orf72 protein, human
  • RNA, Messenger