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Genetic background affects relative nonsense mRNA accumulation in wild-type and upf mutant yeast strains
Bessie Kebaara1, Tara Nazarenus, Rachel Taylor
1School of Biological Sciences, University of Nebraska-Lincoln, Lincoln, NE 68588-0666, USA.
Abstract:
The Saccharomyces cerevisiae nonsense-mediated mRNA decay (NMD) pathway targets mRNAs with premature stop codons and some wild-type mRNAs for accelerated decay. Upf1p, Upf2p and Upf3p are required for NMD. NMD-targeted mRNAs are degraded rapidly in wild-type cells and stabilized in upf1, upf2 or upf3 mutants. We report here that the relative CYH2 pre-mRNA/mRNA accumulation is enhanced in cells derived from a W303 background, compared with a variety of commonly used strains. The enhanced CYH2 pre-mRNA accumulation phenotype results from a larger difference in mRNA half-lives in the W303 strains than two previously used strains. This phenotype can be selected in crosses and is also seen in upf2 and upf3 mutants. These results suggest there are genes that influence the efficiency of NMD and that yeast strains derived from the W303 background may be useful for measurement of abundance and half-lives of low abundance, short-lived NMD substrates.
Insights
Yeast strains from the W303 background show enhanced nonsense-mediated mRNA decay (NMD) pathway activity. This finding suggests specific genes influence NMD efficiency, aiding the study of short-lived mRNA decay.
Area of Science:
- Molecular Biology
- Genetics
- Yeast Biology
Background:
- The nonsense-mediated mRNA decay (NMD) pathway degrades specific mRNAs, including those with premature stop codons.
- Key proteins Upf1p, Upf2p, and Upf3p are essential for NMD function in Saccharomyces cerevisiae.
Purpose of the Study:
- To investigate variations in NMD pathway efficiency across different yeast strains.
- To identify yeast strain backgrounds that may enhance the study of NMD substrates.
Main Methods:
- Comparative analysis of CYH2 pre-mRNA/mRNA accumulation in various Saccharomyces cerevisiae strains.
- Genetic crosses to select for enhanced NMD phenotypes.
- Assessment of mRNA half-lives in different genetic backgrounds.
Main Results:
- Yeast strains from the W303 background exhibit significantly enhanced CYH2 pre-mRNA/mRNA accumulation compared to other common strains.
- This enhanced accumulation is due to a larger difference in mRNA half-lives in W303 strains.
- The phenotype is observable in upf2 and upf3 mutants and can be selected through genetic crosses.
Conclusions:
- The efficiency of the NMD pathway is influenced by specific genes.
- Yeast strains derived from the W303 background are valuable tools for measuring the abundance and half-lives of low-abundance, short-lived NMD substrates.