Impact of Pus1 Pseudouridine Synthase on Specific Decoding Events in Saccharomyces cerevisiae

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.

Medienart:

E-Artikel

Erscheinungsjahr:

2020

Erschienen:

2020

Enthalten in:

Zur Gesamtaufnahme - volume:10

Enthalten in:

Biomolecules - 10(2020), 5 vom: 07. Mai

Sprache:

Englisch

Beteiligte Personen:

Khonsari, Bahar [VerfasserIn]
Klassen, Roland [VerfasserIn]

Links:

Volltext

Themen:

1445-07-4
9014-25-9
Codon
EC 5.4.-
EC 5.4.99.-
Intramolecular Transferases
Journal Article
Misreading
Non-sense suppression
PUS1
Pseudouridine
Pseudouridine synthases
RNA, Transfer
Rapid tRNA decay
Research Support, Non-U.S. Gov't
Saccharomyces cerevisiae Proteins
Sup70-65
TRNA modification
Translation

Anmerkungen:

Date Completed 08.04.2021

Date Revised 08.04.2021

published: Electronic

Citation Status MEDLINE

doi:

10.3390/biom10050729

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

NLM309759609