University of Massachusetts Medical School Faculty Publications
UMMS Affiliation
Department of Microbiology and Physiological Systems
Publication Date
2018-05-24
Document Type
Article Preprint
Disciplines
Amino Acids, Peptides, and Proteins | Biochemistry | Nucleic Acids, Nucleotides, and Nucleosides
Abstract
Nonsense suppressors (NonSups) treat premature termination codon (PTC) disorders by inducing the selection of near cognate tRNAs at the PTC position, allowing readthrough of the PTC and production of full-length protein. Studies of NonSup-induced readthrough of eukaryotic PTCs have been carried out using animals, cells or crude cell extracts. In these studies, NonSups can promote readthrough directly, by binding to components of the protein synthesis machinery, or indirectly, by inhibiting nonsense-mediated mRNA decay or by other mechanisms. Here we utilize a highly-purified in vitro system (Zhang et al., 2016. eLife 5: e13429) to measure exclusively direct NonSup-induced readthrough. Of 17 previously identified NonSups, 13 display direct effects, apparently via at least two different mechanisms. We can monitor such direct effects by single molecule FRET (smFRET). Future smFRET experiments will permit elucidation of the mechanisms by which NonSups stimulate direct readthrough, aiding ongoing efforts to improve the clinical usefulness of NonSups.
Keywords
nonsense suppressors, eukaryotic protein synthesis machinery, single molecule FRET, biochemistry
Rights and Permissions
The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY 4.0 International license.
DOI of Published Version
10.1101/330506
Source
bioRxiv 330506; doi: https://doi.org/10.1101/330506. Link to preprint on bioRxiv service.
Journal/Book/Conference Title
bioRxiv
Repository Citation
Ng, Martin Y.; Zhang, Haibo; Weil, Amy; Singh, Vijay; Jamiolkowski, Ryan M.; Baradaran-Heravi, Alireza; Roberge, Michel; Jacobson, Allan; Welch, Ellen; Goldman, Yale; and Cooperman, Barry S., "A new in vitro assay measuring direct interaction of nonsense suppressors with the eukaryotic protein synthesis machinery" (2018). University of Massachusetts Medical School Faculty Publications. 1501.
https://escholarship.umassmed.edu/faculty_pubs/1501
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.
Included in
Amino Acids, Peptides, and Proteins Commons, Biochemistry Commons, Nucleic Acids, Nucleotides, and Nucleosides Commons