Inactivation of NMD increases viability of sup45 nonsense mutants in Saccharomyces cerevisiae

Svetlana Chabelskaya, Valentina Gryzina, Svetlana Moskalenko, Catherine Le Goff, Galina Zhouravleva

Research output

14 Citations (Scopus)

Abstract

Background: The nonsense-mediated mRNA decay (NMD) pathway promotes the rapid degradation of mRNAs containing premature termination codons (PTCs). In yeast Saccharomyces cerevisiae, the activity of the NMD pathway depends on the recognition of the PTC by the translational machinery. Translation termination factors eRF1 (Sup45) and eRF3 (Sup35) participate not only in the last step of protein synthesis but also in mRNA degradation and translation initiation via interaction with such proteins as Pab1, Upf1, Upf2 and Upf3. Results: In this work we have used previously isolated sup45 mutants of S. cerevisiae to characterize degradation of aberrant mRNA in conditions when translation termination is impaired. We have sequenced his7-1, lys9-A21 and trp1-289 alleles which are frequently used for analysis of nonsense suppression. We have established that sup45 nonsense and missense mutations lead to accumulation of his7-1 mRNA and CYH2 pre-mRNA. Remarkably, deletion of the UPF1 gene suppresses some sup45 phenotypes. In particular, sup45-n upf1δ double mutants were less temperature sensitive, and more resistant to paromomycin than sup45 single mutants. In addition, deletion of either UPF2 or UPF3 restored viability of sup45-n double mutants. Conclusion: This is the first demonstration that sup45 mutations do not only change translation fidelity but also acts by causing a change in mRNA stability.

Original languageEnglish
Article number71
JournalBMC Molecular Biology
Volume8
DOIs
Publication statusPublished - 16 Aug 2007

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Nonsense Mediated mRNA Decay
RNA Stability
Saccharomyces cerevisiae
Nonsense Codon
Paromomycin
RNA Precursors
Gene Deletion
Protein Biosynthesis
Missense Mutation
Proteins
Yeasts
Alleles
Phenotype
Messenger RNA
Mutation
Temperature

Scopus subject areas

  • Molecular Biology

Cite this

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title = "Inactivation of NMD increases viability of sup45 nonsense mutants in Saccharomyces cerevisiae",
abstract = "Background: The nonsense-mediated mRNA decay (NMD) pathway promotes the rapid degradation of mRNAs containing premature termination codons (PTCs). In yeast Saccharomyces cerevisiae, the activity of the NMD pathway depends on the recognition of the PTC by the translational machinery. Translation termination factors eRF1 (Sup45) and eRF3 (Sup35) participate not only in the last step of protein synthesis but also in mRNA degradation and translation initiation via interaction with such proteins as Pab1, Upf1, Upf2 and Upf3. Results: In this work we have used previously isolated sup45 mutants of S. cerevisiae to characterize degradation of aberrant mRNA in conditions when translation termination is impaired. We have sequenced his7-1, lys9-A21 and trp1-289 alleles which are frequently used for analysis of nonsense suppression. We have established that sup45 nonsense and missense mutations lead to accumulation of his7-1 mRNA and CYH2 pre-mRNA. Remarkably, deletion of the UPF1 gene suppresses some sup45 phenotypes. In particular, sup45-n upf1δ double mutants were less temperature sensitive, and more resistant to paromomycin than sup45 single mutants. In addition, deletion of either UPF2 or UPF3 restored viability of sup45-n double mutants. Conclusion: This is the first demonstration that sup45 mutations do not only change translation fidelity but also acts by causing a change in mRNA stability.",
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Inactivation of NMD increases viability of sup45 nonsense mutants in Saccharomyces cerevisiae. / Chabelskaya, Svetlana; Gryzina, Valentina; Moskalenko, Svetlana; Le Goff, Catherine; Zhouravleva, Galina.

In: BMC Molecular Biology, Vol. 8, 71, 16.08.2007.

Research output

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T1 - Inactivation of NMD increases viability of sup45 nonsense mutants in Saccharomyces cerevisiae

AU - Chabelskaya, Svetlana

AU - Gryzina, Valentina

AU - Moskalenko, Svetlana

AU - Le Goff, Catherine

AU - Zhouravleva, Galina

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AB - Background: The nonsense-mediated mRNA decay (NMD) pathway promotes the rapid degradation of mRNAs containing premature termination codons (PTCs). In yeast Saccharomyces cerevisiae, the activity of the NMD pathway depends on the recognition of the PTC by the translational machinery. Translation termination factors eRF1 (Sup45) and eRF3 (Sup35) participate not only in the last step of protein synthesis but also in mRNA degradation and translation initiation via interaction with such proteins as Pab1, Upf1, Upf2 and Upf3. Results: In this work we have used previously isolated sup45 mutants of S. cerevisiae to characterize degradation of aberrant mRNA in conditions when translation termination is impaired. We have sequenced his7-1, lys9-A21 and trp1-289 alleles which are frequently used for analysis of nonsense suppression. We have established that sup45 nonsense and missense mutations lead to accumulation of his7-1 mRNA and CYH2 pre-mRNA. Remarkably, deletion of the UPF1 gene suppresses some sup45 phenotypes. In particular, sup45-n upf1δ double mutants were less temperature sensitive, and more resistant to paromomycin than sup45 single mutants. In addition, deletion of either UPF2 or UPF3 restored viability of sup45-n double mutants. Conclusion: This is the first demonstration that sup45 mutations do not only change translation fidelity but also acts by causing a change in mRNA stability.

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