Incorporation of pseudouridine into mRNA enhances translation by diminishing PKR activation
Top Cited Papers
Open Access
- 10 May 2010
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 38 (17) , 5884-5892
- https://doi.org/10.1093/nar/gkq347
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.Keywords
This publication has 56 references indexed in Scilit:
- Influenza B Virus Ribonucleoprotein Is a Potent Activator of the Antiviral Kinase PKRPLoS Pathogens, 2009
- Incorporation of Pseudouridine Into mRNA Yields Superior Nonimmunogenic Vector With Increased Translational Capacity and Biological StabilityMolecular Therapy, 2008
- Nucleoside modifications modulate activation of the protein kinase PKR in an RNA structure-specific mannerRNA, 2008
- 5'-Triphosphate RNA Is the Ligand for RIG-IScience, 2006
- Controlling activation of the RNA-dependent protein kinase by siRNAs using site-specific chemical modificationNucleic Acids Research, 2006
- Suppression of RNA Recognition by Toll-like Receptors: The Impact of Nucleoside Modification and the Evolutionary Origin of RNAImmunity, 2005
- Activation of double-stranded RNA-dependent kinase (dsl) by the TAR region of HIV-1 mRNA: A novel translational control mechanismCell, 1989
- Activation of human and mouse 2‐5A synthetases and mouse protein P1 kinase by nucleic acidsFEBS Letters, 1981
- Differential effects of various double-stranded RNAs on protein synthesis in rabbit reticulocyte lysatesBiochemistry, 1978
- Complexing ability and coding properties of synthetic polynucleotidesJournal of Molecular Biology, 1964