Impact of Pus1 Pseudouridine Synthase on Specific Decoding Events in <i>Saccharomyces cerevisiae</i>
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 <i>PUS1</i> deletion on the essential decoding of CAG (Gln) codons by tRNA<sup>Gln</sup>...
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MDPI AG
2020-05-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/10/5/729 |
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| author | Bahar Khonsari Roland Klassen |
| author_facet | Bahar Khonsari Roland Klassen |
| author_sort | Bahar Khonsari |
| collection | DOAJ |
| description | 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 <i>PUS1</i> deletion on the essential decoding of CAG (Gln) codons by tRNA<sup>Gln</sup>CUG in yeast. Synthetic lethality was observed upon combining the modification defect with destabilized variants of tRNA<sup>Gln</sup>CUG, pointing to a severe CAG-decoding defect of the hypomodified tRNA. In addition, we demonstrated that misreading of UAG stop codons by a tRNA<sup>Gln</sup>CUG variant is positively affected by Pus1. Genetic approaches further indicated that mildly elevated temperature decreases the decoding efficiency of CAG and UAG via destabilized tRNA<sup>Gln</sup>CAG variants. We also determined the misreading of CGC (Arg) codons by tRNA<sup>His</sup>GUG, where the CGC decoder tRNA<sup>Arg</sup>ICG contains Pus1-dependent pseudouridine, but not the mistranslating tRNA<sup>His</sup>. We found that the absence of Pus1 increased CGC misreading by tRNA<sup>His</sup>, 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 <i>pus1</i> mutants and strains carrying destabilized tRNA<sup>Gln</sup>CAG were suppressible by additional deletion of the rapid tRNA decay (RTD)-relevant <i>MET22</i>, suggesting the involvement of RTD-mediated tRNA destabilization. |
| first_indexed | 2024-03-10T19:59:15Z |
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| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-10T19:59:15Z |
| publishDate | 2020-05-01 |
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| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-4aff0823c52c4c82bcc8638aa9b009fd2023-11-19T23:42:26ZengMDPI AGBiomolecules2218-273X2020-05-0110572910.3390/biom10050729Impact of Pus1 Pseudouridine Synthase on Specific Decoding Events in <i>Saccharomyces cerevisiae</i>Bahar Khonsari0Roland Klassen1Institut für Biologie, Fachgebiet Mikrobiologie, Universität Kassel, Heinrich-Plett-Str. 40, D-34132 Kassel, GermanyInstitut für Biologie, Fachgebiet Mikrobiologie, Universität Kassel, Heinrich-Plett-Str. 40, D-34132 Kassel, GermanyPus1-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 <i>PUS1</i> deletion on the essential decoding of CAG (Gln) codons by tRNA<sup>Gln</sup>CUG in yeast. Synthetic lethality was observed upon combining the modification defect with destabilized variants of tRNA<sup>Gln</sup>CUG, pointing to a severe CAG-decoding defect of the hypomodified tRNA. In addition, we demonstrated that misreading of UAG stop codons by a tRNA<sup>Gln</sup>CUG variant is positively affected by Pus1. Genetic approaches further indicated that mildly elevated temperature decreases the decoding efficiency of CAG and UAG via destabilized tRNA<sup>Gln</sup>CAG variants. We also determined the misreading of CGC (Arg) codons by tRNA<sup>His</sup>GUG, where the CGC decoder tRNA<sup>Arg</sup>ICG contains Pus1-dependent pseudouridine, but not the mistranslating tRNA<sup>His</sup>. We found that the absence of Pus1 increased CGC misreading by tRNA<sup>His</sup>, 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 <i>pus1</i> mutants and strains carrying destabilized tRNA<sup>Gln</sup>CAG were suppressible by additional deletion of the rapid tRNA decay (RTD)-relevant <i>MET22</i>, suggesting the involvement of RTD-mediated tRNA destabilization.https://www.mdpi.com/2218-273X/10/5/729tRNA modification<i>PUS1</i>pseudouridinetranslationnon-sense suppression<i>sup70-65</i> |
| spellingShingle | Bahar Khonsari Roland Klassen Impact of Pus1 Pseudouridine Synthase on Specific Decoding Events in <i>Saccharomyces cerevisiae</i> Biomolecules tRNA modification <i>PUS1</i> pseudouridine translation non-sense suppression <i>sup70-65</i> |
| title | Impact of Pus1 Pseudouridine Synthase on Specific Decoding Events in <i>Saccharomyces cerevisiae</i> |
| title_full | Impact of Pus1 Pseudouridine Synthase on Specific Decoding Events in <i>Saccharomyces cerevisiae</i> |
| title_fullStr | Impact of Pus1 Pseudouridine Synthase on Specific Decoding Events in <i>Saccharomyces cerevisiae</i> |
| title_full_unstemmed | Impact of Pus1 Pseudouridine Synthase on Specific Decoding Events in <i>Saccharomyces cerevisiae</i> |
| title_short | Impact of Pus1 Pseudouridine Synthase on Specific Decoding Events in <i>Saccharomyces cerevisiae</i> |
| title_sort | impact of pus1 pseudouridine synthase on specific decoding events in i saccharomyces cerevisiae i |
| topic | tRNA modification <i>PUS1</i> pseudouridine translation non-sense suppression <i>sup70-65</i> |
| url | https://www.mdpi.com/2218-273X/10/5/729 |
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