Arginine-rich C9ORF72 ALS proteins stall ribosomes in a manner distinct from a canonical ribosome-associated quality control substrate
- Author(s)
- Kriachkov, V; Ormsby, AR; Kusnadi, EP; McWilliam, HEG; Mintern, JD; Amarasinghe, SL; Ritchie, ME; Furic, L; Hatters, DM;
- Details
- Publication Year 2023-01,Volume 299,Issue #1,Page 102774
- Journal Title
- Journal of Biological Chemistry
- Publication Type
- Research article
- Abstract
- Hexanucleotide expansion mutations in C9ORF72 are a frequent cause of amyotrophic lateral sclerosis. We previously reported that long arginine-rich dipeptide repeats (DPRs), mimicking abnormal proteins expressed from the hexanucleotide expansion, caused translation stalling when expressed in cell culture models. Whether this stalling provides a mechanism of pathogenicity remains to be determined. Here, we explored the molecular features of DPR-induced stalling and examined whether known mechanisms such as ribosome quality control (RQC) regulate translation elongation on sequences that encode arginine-rich DPRs. We demonstrate that arginine-rich DPRs lead to stalling in a length-dependent manner, with lengths longer than 40 repeats invoking severe translation arrest. Mutational screening of 40xGly-Xxx DPRs shows that stalling is most pronounced when Xxx is a charged amino acid (Arg, Lys, Glu, or Asp). Through a genome-wide knockout screen, we find that genes regulating stalling on polyadenosine mRNA coding for poly-Lys, a canonical RQC substrate, act differently in the case of arginine-rich DPRs. Indeed, these findings point to a limited scope for natural regulatory responses to resolve the arginine-rich DPR stalls, even though the stalls may be sensed, as evidenced by an upregulation of RQC gene expression. These findings therefore implicate arginine-rich DPR-mediated stalled ribosomes as a source of stress and toxicity and may be a crucial component in pathomechanisms.
- Publisher
- Elsevier
- Keywords
- Humans; *Amyotrophic Lateral Sclerosis/genetics/physiopathology; Arginine/metabolism; C9orf72 Protein/genetics/metabolism; Dipeptides/chemistry; Ribosomes/genetics/metabolism; Gene Knockout Techniques; Mutation; Up-Regulation; C9orf72 als; RNA-protein interaction; Rqc; arginine-rich dipeptide repeats; neurodegenerative disease; protein misfolding; ribosome function; ribosome stalling; translation
- Department(s)
- Laboratory Research
- PubMed ID
- 36481270
- Publisher's Version
- https://doi.org/10.1016/j.jbc.2022.102774
- Terms of Use/Rights Notice
- Refer to copyright notice on published article.
Creation Date: 2023-05-30 07:27:56
Last Modified: 2024-07-09 06:24:24