Incorporation of pseudouridine into mRNA enhances translation by diminishing PKR activation
Year: 2010
Type: article
Abstract: Previous studies have shown that the translation level of in vitro transcribed messenger RNA (mRNA) is enhanced when its uridines are replaced with pseudouridines; however, the reason for this enhancement has not been identified. Here, we demonstrate that in vitro transcripts containing uridine activate RNA-dependent protein kinase (PKR), which then phosphorylates translation initiation factor 2-alpha (eIF-2α), and inhibits translation. In contrast, in vitro transcribed mRNAs containing pseudouridine activate PKR to a lesser degree, and translation of pseudouridine-containing mRNAs is not repressed. RNA pull-down assays demonstrate that mRNA containing uridine is bound by PKR more efficiently than mRNA with pseudouridine. Finally, the role of PKR is validated by showing that pseudouridine- and uridine-containing RNAs were translated equally in PKR knockout cells. These results indicate that the enhanced translation of mRNAs containing pseudouridine, compared to those containing uridine, is mediated by decreased activation of PKR.
Source: Nucleic Acids Research
DOI: 10.1093/nar/gkq347
Institutions: Pennsylvania State University, University of Pennsylvania
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,992
Funders: Cleveland Clinic, Cleveland Clinic Foundation, National Institutes of Health, National Heart, Lung, and Blood Institute, National Institute of General Medical Sciences, National Institute of Allergy and Infectious Diseases, National Institute of Diabetes and Digestive and Kidney Diseases, National Institute of Dental and Craniofacial Research, National Center for Research Resources
Awards: Gene therapy training: CF and genetic diseases, G2213370879, Training Program in Cell and Molecular Biology, G3879651179, RNA delivery for dendritic cell HIV antigen presentation, G4957090920, G572917779, G618623535, Oral Delivery of RNA Encoded Antigen, Training Grant in Comparative Medical & Molecular Genetics, G972951537
OpenAlex ID: W2156788486