Glutamine missense suppressor transfer RNAs inhibit polyglutamine aggregation
Huntington’s disease (HD) is caused by polyglutamine (polyQ) repeat expansions in the huntingtin gene. HD-causative polyQ alleles lead to protein aggregation, which is a prerequisite for disease. Translation fidelity modifies protein aggregation, and several studies suggest that mutating one or two...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-03-01
|
| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253124003299 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832592819562217472 |
|---|---|
| author | Rasangi Tennakoon Teija M.I. Bily Farah Hasan Sunidhi Syal Aaron Voigt Tugce B. Balci Kyle S. Hoffman Patrick O’Donoghue |
| author_facet | Rasangi Tennakoon Teija M.I. Bily Farah Hasan Sunidhi Syal Aaron Voigt Tugce B. Balci Kyle S. Hoffman Patrick O’Donoghue |
| author_sort | Rasangi Tennakoon |
| collection | DOAJ |
| description | Huntington’s disease (HD) is caused by polyglutamine (polyQ) repeat expansions in the huntingtin gene. HD-causative polyQ alleles lead to protein aggregation, which is a prerequisite for disease. Translation fidelity modifies protein aggregation, and several studies suggest that mutating one or two glutamine (Gln) residues in polyQ reduces aggregation. Thus, we hypothesized that missense suppression of Gln codons with other amino acids will reduce polyQ aggregate formation in cells. In neuroblastoma cells, we assessed tRNA variants that misread Gln codons with serine (tRNASerC/UUG) or alanine (tRNAAlaC/UUG). The tRNAs with the CUG anticodon were more effective at suppressing the CAG repeats in polyQ, and serine and alanine mis-incorporation had differential impacts on polyQ. The expression of tRNASerCUG reduced polyQ protein production as well as both soluble and insoluble aggregate formation. In contrast, cells expressing tRNAAlaCUG selectively decreased insoluble polyQ aggregate formation by 2-fold. Mass spectrometry confirmed Ala mis-incorporation at an average level of ∼20% per Gln codon. Cells expressing the missense suppressor tRNAs showed no cytotoxic effects and no defects in growth or global protein synthesis levels. Our findings demonstrate that tRNA-dependent missense suppression of Gln codons is well tolerated in mammalian cells and significantly reduces polyQ levels and aggregates that cause HD. |
| format | Article |
| id | doaj-art-d4938172ff0c4ddf935181b8f0c1e460 |
| institution | Kabale University |
| issn | 2162-2531 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj-art-d4938172ff0c4ddf935181b8f0c1e4602025-01-21T04:13:04ZengElsevierMolecular Therapy: Nucleic Acids2162-25312025-03-01361102442Glutamine missense suppressor transfer RNAs inhibit polyglutamine aggregationRasangi Tennakoon0Teija M.I. Bily1Farah Hasan2Sunidhi Syal3Aaron Voigt4Tugce B. Balci5Kyle S. Hoffman6Patrick O’Donoghue7Department of Biochemistry, The University of Western Ontario, London, ON N6A 5C1, CanadaDepartment of Biochemistry, The University of Western Ontario, London, ON N6A 5C1, CanadaDepartment of Biochemistry, The University of Western Ontario, London, ON N6A 5C1, CanadaDepartment of Biochemistry, The University of Western Ontario, London, ON N6A 5C1, CanadaDepartment of Neurology, RWTH Aachen, 52062 Aachen, GermanyDepartment of Paediatrics, The University of Western Ontario, London, ON N6A 5C1, CanadaBioinformatic Solutions, Inc, Waterloo, ON N2L 3K8, CanadaDepartment of Biochemistry, The University of Western Ontario, London, ON N6A 5C1, Canada; Department of Chemistry, The University of Western Ontario, London, ON N6A 5C1, Canada; Corresponding author: Patrick O’Donoghue, Department of Biochemistry, The University of Western Ontario, London, ON N6A 5C1, CanadaHuntington’s disease (HD) is caused by polyglutamine (polyQ) repeat expansions in the huntingtin gene. HD-causative polyQ alleles lead to protein aggregation, which is a prerequisite for disease. Translation fidelity modifies protein aggregation, and several studies suggest that mutating one or two glutamine (Gln) residues in polyQ reduces aggregation. Thus, we hypothesized that missense suppression of Gln codons with other amino acids will reduce polyQ aggregate formation in cells. In neuroblastoma cells, we assessed tRNA variants that misread Gln codons with serine (tRNASerC/UUG) or alanine (tRNAAlaC/UUG). The tRNAs with the CUG anticodon were more effective at suppressing the CAG repeats in polyQ, and serine and alanine mis-incorporation had differential impacts on polyQ. The expression of tRNASerCUG reduced polyQ protein production as well as both soluble and insoluble aggregate formation. In contrast, cells expressing tRNAAlaCUG selectively decreased insoluble polyQ aggregate formation by 2-fold. Mass spectrometry confirmed Ala mis-incorporation at an average level of ∼20% per Gln codon. Cells expressing the missense suppressor tRNAs showed no cytotoxic effects and no defects in growth or global protein synthesis levels. Our findings demonstrate that tRNA-dependent missense suppression of Gln codons is well tolerated in mammalian cells and significantly reduces polyQ levels and aggregates that cause HD.http://www.sciencedirect.com/science/article/pii/S2162253124003299MT: Non-coding RNAsHuntington's diseaseHDgene therapymistranslationmissense suppression |
| spellingShingle | Rasangi Tennakoon Teija M.I. Bily Farah Hasan Sunidhi Syal Aaron Voigt Tugce B. Balci Kyle S. Hoffman Patrick O’Donoghue Glutamine missense suppressor transfer RNAs inhibit polyglutamine aggregation Molecular Therapy: Nucleic Acids MT: Non-coding RNAs Huntington's disease HD gene therapy mistranslation missense suppression |
| title | Glutamine missense suppressor transfer RNAs inhibit polyglutamine aggregation |
| title_full | Glutamine missense suppressor transfer RNAs inhibit polyglutamine aggregation |
| title_fullStr | Glutamine missense suppressor transfer RNAs inhibit polyglutamine aggregation |
| title_full_unstemmed | Glutamine missense suppressor transfer RNAs inhibit polyglutamine aggregation |
| title_short | Glutamine missense suppressor transfer RNAs inhibit polyglutamine aggregation |
| title_sort | glutamine missense suppressor transfer rnas inhibit polyglutamine aggregation |
| topic | MT: Non-coding RNAs Huntington's disease HD gene therapy mistranslation missense suppression |
| url | http://www.sciencedirect.com/science/article/pii/S2162253124003299 |
| work_keys_str_mv | AT rasangitennakoon glutaminemissensesuppressortransferrnasinhibitpolyglutamineaggregation AT teijamibily glutaminemissensesuppressortransferrnasinhibitpolyglutamineaggregation AT farahhasan glutaminemissensesuppressortransferrnasinhibitpolyglutamineaggregation AT sunidhisyal glutaminemissensesuppressortransferrnasinhibitpolyglutamineaggregation AT aaronvoigt glutaminemissensesuppressortransferrnasinhibitpolyglutamineaggregation AT tugcebbalci glutaminemissensesuppressortransferrnasinhibitpolyglutamineaggregation AT kyleshoffman glutaminemissensesuppressortransferrnasinhibitpolyglutamineaggregation AT patrickodonoghue glutaminemissensesuppressortransferrnasinhibitpolyglutamineaggregation |