Single-molecule imaging reveals distinct elongation and frameshifting dynamics between frames of expanded RNA repeats in C9ORF72-ALS/FTD
Abstract C9ORF72 hexanucleotide repeat expansion is the most common genetic cause of both amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). One pathogenic mechanism is the accumulation of toxic dipeptide repeat (DPR) proteins like poly-GA, GP and GR, produced by the noncanonical...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-09-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-023-41339-x |
| _version_ | 1797557890068774912 |
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| author | Malgorzata J. Latallo Shaopeng Wang Daoyuan Dong Blake Nelson Nathan M. Livingston Rong Wu Ning Zhao Timothy J. Stasevich Michael C. Bassik Shuying Sun Bin Wu |
| author_facet | Malgorzata J. Latallo Shaopeng Wang Daoyuan Dong Blake Nelson Nathan M. Livingston Rong Wu Ning Zhao Timothy J. Stasevich Michael C. Bassik Shuying Sun Bin Wu |
| author_sort | Malgorzata J. Latallo |
| collection | DOAJ |
| description | Abstract C9ORF72 hexanucleotide repeat expansion is the most common genetic cause of both amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). One pathogenic mechanism is the accumulation of toxic dipeptide repeat (DPR) proteins like poly-GA, GP and GR, produced by the noncanonical translation of the expanded RNA repeats. However, how different DPRs are synthesized remains elusive. Here, we use single-molecule imaging techniques to directly measure the translation dynamics of different DPRs. Besides initiation, translation elongation rates vary drastically between different frames, with GP slower than GA and GR the slowest. We directly visualize frameshift events using a two-color single-molecule translation assay. The repeat expansion enhances frameshifting, but the overall frequency is low. There is a higher chance of GR-to-GA shift than in the reversed direction. Finally, the ribosome-associated protein quality control (RQC) factors ZNF598 and Pelota modulate the translation dynamics, and the repeat RNA sequence is important for invoking the RQC pathway. This study reveals that multiple translation steps modulate the final DPR production. Understanding repeat RNA translation is critically important to decipher the DPR-mediated pathogenesis and identify potential therapeutic targets in C9ORF72-ALS/FTD. |
| first_indexed | 2024-03-10T17:22:43Z |
| format | Article |
| id | doaj.art-ab9f2ebcd16e4163807ff2dcc11f4974 |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-03-10T17:22:43Z |
| publishDate | 2023-09-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-ab9f2ebcd16e4163807ff2dcc11f49742023-11-20T10:17:00ZengNature PortfolioNature Communications2041-17232023-09-0114111810.1038/s41467-023-41339-xSingle-molecule imaging reveals distinct elongation and frameshifting dynamics between frames of expanded RNA repeats in C9ORF72-ALS/FTDMalgorzata J. Latallo0Shaopeng Wang1Daoyuan Dong2Blake Nelson3Nathan M. Livingston4Rong Wu5Ning Zhao6Timothy J. Stasevich7Michael C. Bassik8Shuying Sun9Bin Wu10Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of MedicineDepartment of Biophysics and Biophysical Chemistry, Johns Hopkins University School of MedicineDepartment of Physiology, Johns Hopkins University School of MedicineDepartment of Biophysics and Biophysical Chemistry, Johns Hopkins University School of MedicineDepartment of Biophysics and Biophysical Chemistry, Johns Hopkins University School of MedicineDepartment of Physiology, Johns Hopkins University School of MedicineDepartment of Biochemistry and Molecular Genetics, University of Colorado-Anschutz Medical CampusDepartment of Biochemistry and Molecular Genetics, University of Colorado-Anschutz Medical CampusDepartment of Genetics, Stanford University School of MedicineCenter for Cell Dynamics, Johns Hopkins University School of MedicineDepartment of Biophysics and Biophysical Chemistry, Johns Hopkins University School of MedicineAbstract C9ORF72 hexanucleotide repeat expansion is the most common genetic cause of both amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). One pathogenic mechanism is the accumulation of toxic dipeptide repeat (DPR) proteins like poly-GA, GP and GR, produced by the noncanonical translation of the expanded RNA repeats. However, how different DPRs are synthesized remains elusive. Here, we use single-molecule imaging techniques to directly measure the translation dynamics of different DPRs. Besides initiation, translation elongation rates vary drastically between different frames, with GP slower than GA and GR the slowest. We directly visualize frameshift events using a two-color single-molecule translation assay. The repeat expansion enhances frameshifting, but the overall frequency is low. There is a higher chance of GR-to-GA shift than in the reversed direction. Finally, the ribosome-associated protein quality control (RQC) factors ZNF598 and Pelota modulate the translation dynamics, and the repeat RNA sequence is important for invoking the RQC pathway. This study reveals that multiple translation steps modulate the final DPR production. Understanding repeat RNA translation is critically important to decipher the DPR-mediated pathogenesis and identify potential therapeutic targets in C9ORF72-ALS/FTD.https://doi.org/10.1038/s41467-023-41339-x |
| spellingShingle | Malgorzata J. Latallo Shaopeng Wang Daoyuan Dong Blake Nelson Nathan M. Livingston Rong Wu Ning Zhao Timothy J. Stasevich Michael C. Bassik Shuying Sun Bin Wu Single-molecule imaging reveals distinct elongation and frameshifting dynamics between frames of expanded RNA repeats in C9ORF72-ALS/FTD Nature Communications |
| title | Single-molecule imaging reveals distinct elongation and frameshifting dynamics between frames of expanded RNA repeats in C9ORF72-ALS/FTD |
| title_full | Single-molecule imaging reveals distinct elongation and frameshifting dynamics between frames of expanded RNA repeats in C9ORF72-ALS/FTD |
| title_fullStr | Single-molecule imaging reveals distinct elongation and frameshifting dynamics between frames of expanded RNA repeats in C9ORF72-ALS/FTD |
| title_full_unstemmed | Single-molecule imaging reveals distinct elongation and frameshifting dynamics between frames of expanded RNA repeats in C9ORF72-ALS/FTD |
| title_short | Single-molecule imaging reveals distinct elongation and frameshifting dynamics between frames of expanded RNA repeats in C9ORF72-ALS/FTD |
| title_sort | single molecule imaging reveals distinct elongation and frameshifting dynamics between frames of expanded rna repeats in c9orf72 als ftd |
| url | https://doi.org/10.1038/s41467-023-41339-x |
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