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Mutationally altered 3' ends of yeast CYC1 mRNA affect transcript stability and translational efficiency
Journal of Molecular Biology
|July 25, 1984
Summary
Genetic alterations in yeast Saccharomyces cerevisiae can restore CYC1 mRNA stability and function. These changes involve chromosomal rearrangements, local mutations, or suppressors, impacting transcription termination and mRNA processing.
Area of Science:
- Molecular Biology
- Yeast Genetics
- Gene Regulation
Background:
- The cyc1-512 mutant in Saccharomyces cerevisiae has a deletion in the 3' non-coding region of the CYC1 gene, leading to unstable and elongated CYC1 messenger RNA (mRNA).
- Previous studies identified three classes of genetic events—chromosomal rearrangements, local mutations, and unlinked suppressors—that can restore function to the cyc1-512 mutant.
Purpose of the Study:
- To investigate the molecular mechanisms by which different genetic events restore function to the cyc1-512 yeast mutant.
- To analyze the impact of altered 3' non-coding sequences on CYC1 mRNA stability, translational efficiency, and transcription termination.
Main Methods:
- Analysis of revertants of the cyc1-512 mutant, including those with chromosomal rearrangements and local genetic changes.
- Sequencing of DNA to identify specific genetic alterations and their locations relative to the CYC1 locus.
- Characterization of CYC1 mRNA transcripts, including length, stability, and termination sites.
- Identification and characterization of extragenic suppressors affecting termination and mRNA stability.
Main Results:
- Chromosomal rearrangements 3' to the CYC1 locus create new 3' non-coding regions, enhancing mRNA stability with varied translational effects.
- Local genetic changes in revertants generated stronger transcription terminators, with some exhibiting discrete termination and others heterogeneous termination.
- Extragenic suppressors were identified that either enhance termination at weak sites or increase the stability of elongated CYC1 mRNA.
Conclusions:
- The 3' non-coding region plays a critical role in CYC1 mRNA stability and translational efficiency in yeast.
- Transcription termination in yeast can be modulated by various genetic events, including rearrangements and local sequence changes.
- Findings support the hypothesis that transcription termination and polyadenylation are coupled processes in yeast, unlike in many higher eukaryotes.
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