Date
2020-05-07Metadata
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Aufsatz
Impact of Pus1 Pseudouridine Synthase on Specific Decoding Events in Saccharomyces cerevisiae
Abstract
Pus1-dependent pseudouridylation occurs in many tRNAs and at multiple positions, yet the functional impact of this modification is incompletely understood. We analyzed the consequences of PUS1 deletion on the essential decoding of CAG (Gln) codons by tRNAGlnCUG in yeast. Synthetic lethality was observed upon combining the modification defect with destabilized variants of tRNAGlnCUG, pointing to a severe CAG-decoding defect of the hypomodified tRNA. In addition, we demonstrated that misreading of UAG stop codons by a tRNAGlnCUG variant is positively affected by Pus1. Genetic approaches further indicated that mildly elevated temperature decreases the decoding efficiency of CAG and UAG via destabilized tRNAGlnCAG variants. We also determined the misreading of CGC (Arg) codons by tRNAHisGUG, where the CGC decoder tRNAArgICG contains Pus1-dependent pseudouridine, but not the mistranslating tRNAHis. We found that the absence of Pus1 increased CGC misreading by tRNAHis, demonstrating a positive role of the modification in the competition against non-synonymous near-cognate tRNA. Part of the in vivo decoding defects and phenotypes in pus1 mutants and strains carrying destabilized tRNAGlnCAG were suppressible by additional deletion of the rapid tRNA decay (RTD)-relevant MET22, suggesting the involvement of RTD-mediated tRNA destabilization.
Citation
In: Biomolecules Volume 10 / Issue 5 (2020-05-07) , S. 729 ; EISSN 2218-273XSponsorship
Gefördert durch den Publikationsfonds der Universität KasselCitation
@article{doi:10.17170/kobra-202006291376,
author={Khonsari, Bahar and Klassen, Roland},
title={Impact of Pus1 Pseudouridine Synthase on Specific Decoding Events in Saccharomyces cerevisiae},
journal={Biomolecules},
year={2020}
}
0500 Oax 0501 Text $btxt$2rdacontent 0502 Computermedien $bc$2rdacarrier 1100 2020$n2020 1500 1/eng 2050 ##0##http://hdl.handle.net/123456789/11611 3000 Khonsari, Bahar 3010 Klassen, Roland 4000 Impact of Pus1 Pseudouridine Synthase on Specific Decoding Events in Saccharomyces cerevisiae / Khonsari, Bahar 4030 4060 Online-Ressource 4085 ##0##=u http://nbn-resolving.de/http://hdl.handle.net/123456789/11611=x R 4204 \$dAufsatz 4170 5550 {{Transfer-RNS}} 5550 {{Pseudouridin}} 7136 ##0##http://hdl.handle.net/123456789/11611
2020-06-29T15:32:58Z 2020-06-29T15:32:58Z 2020-05-07 doi:10.17170/kobra-202006291376 http://hdl.handle.net/123456789/11611 Gefördert durch den Publikationsfonds der Universität Kassel eng Namensnennung 4.0 International http://creativecommons.org/licenses/by/4.0/ tRNA modification PUS1 pseudouridine translation non-sense suppression sup70-65 misreading rapid tRNA decay 570 Impact of Pus1 Pseudouridine Synthase on Specific Decoding Events in Saccharomyces cerevisiae Aufsatz Pus1-dependent pseudouridylation occurs in many tRNAs and at multiple positions, yet the functional impact of this modification is incompletely understood. We analyzed the consequences of PUS1 deletion on the essential decoding of CAG (Gln) codons by tRNAGlnCUG in yeast. Synthetic lethality was observed upon combining the modification defect with destabilized variants of tRNAGlnCUG, pointing to a severe CAG-decoding defect of the hypomodified tRNA. In addition, we demonstrated that misreading of UAG stop codons by a tRNAGlnCUG variant is positively affected by Pus1. Genetic approaches further indicated that mildly elevated temperature decreases the decoding efficiency of CAG and UAG via destabilized tRNAGlnCAG variants. We also determined the misreading of CGC (Arg) codons by tRNAHisGUG, where the CGC decoder tRNAArgICG contains Pus1-dependent pseudouridine, but not the mistranslating tRNAHis. We found that the absence of Pus1 increased CGC misreading by tRNAHis, demonstrating a positive role of the modification in the competition against non-synonymous near-cognate tRNA. Part of the in vivo decoding defects and phenotypes in pus1 mutants and strains carrying destabilized tRNAGlnCAG were suppressible by additional deletion of the rapid tRNA decay (RTD)-relevant MET22, suggesting the involvement of RTD-mediated tRNA destabilization. open access Khonsari, Bahar Klassen, Roland doi:10.3390/biom10050729 Transfer-RNS Pseudouridin publishedVersion EISSN 2218-273X Issue 5 Biomolecules 729 Volume 10 false
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