PXD044322

PXD044322 is an original dataset announced via ProteomeXchange.

Title	Efficient Suppression of Premature Termination Codons by Engineered Pyrrolysine tRNAs
Description	Mutations that introduce premature termination codons (PTCs) within protein-coding genes are associated with incurable and severe genetic diseases. Many PTC-associated disorders are life-threatening and have no approved medical treatment options. Suppressor transfer RNAs (sup-tRNAs) with the capacity to promote translational readthrough of PTCs represent a promising therapeutic strategy to treat these conditions; however, developing novel sup-tRNAs with high efficiency and specificity often requires extensive engineering and screening. Moreover, these efforts are not always successful at producing more efficient sup-tRNAs. Here we show that the pyrrolysine tRNA (tRNAPyl), which naturally translates the UAG stop codon, offers an attractive scaffold for developing potent sup-tRNAs that restore protein synthesis from PTC-containing genes. We created a series of rationally designed Pyrrolysine tRNA Scaffold Suppressor-tRNAs (PASS-tRNAs) that are substrates of bacterial and human alanyl-tRNA synthetase. Using a PTC-containing reporter gene, PASS-tRNAs restore protein synthesis to wild-type levels in bacterial cells. In human cells, PASS-tRNAs display robust and consistent translation activity in two distinct PTC reporter genes with high codon specificity and no observed cytotoxic effects. Collectively, these results unveil a new class of highly efficient sup-tRNAs with great potential for tRNA-based therapeutics.
HostingRepository	PRIDE
AnnounceDate	2025-02-20
AnnouncementXML	Submission_2025-02-20_07:20:40.080.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Jeremy Balsbaugh
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationList	acetylated residue; monohydroxylated residue; deamidated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Eclipse

Revision	Datetime	Status	ChangeLog Entry
0	2023-08-03 07:59:14	ID requested
⏵ 1	2025-02-20 07:20:40	announced

10.1093/nar/gkae1048;

Awawdeh A, Tapia A, Alshawi SA, Dawodu O, Gaier SA, Specht C, Beaudoin JD, Tharp JM, Vargas-Rodriguez O, Efficient suppression of premature termination codons with alanine by engineered chimeric pyrrolysine tRNAs. Nucleic Acids Res, 52(22):14244-14259(2024) [pubmed]

submitter keyword: proteomics,mutations, premature termination codons, tRNA, bacteria, GFP

Jeremy Balsbaugh
contact affiliation	Proteomics & Metabolomics Facility, University of Connecticut
contact email	jeremy.balsbaugh@uconn.edu
lab head
Jeremy Balsbaugh
contact affiliation	University of Connecticut
contact email	jeremy.balsbaugh@uconn.edu
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2025/02/PXD044322

PRIDE project URI

• PRIDE
  1. PXD044322
     1. Label: PRIDE project
     2. Name: Efficient Suppression of Premature Termination Codons by Engineered Pyrrolysine tRNAs