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Single-Molecule mRNA Decay Measurements Reveal Promoter- Regulated mRNA Stability in Yeast
- Trcek, Tatjana, Larson, Daniel R., Moldón, Alberto, Query, Charles C., Singer, Robert H.
- Cell 2011 v.147 no.7 pp. 1484-1497
- complement, fluorescence in situ hybridization, genes, messenger RNA, mitosis, nucleotide sequences, yeasts
- Messenger RNA decay measurements are typically performed on a population of cells. However, this approach cannot reveal sufficient complexity to provide information on mechanisms that may regulate mRNA degradation, possibly on short timescales. To address this deficiency, we measured cell cycle-regulated decay in single yeast cells using single-molecule FISH. We found that two genes responsible for mitotic progression, SWI5 and CLB2, exhibit a mitosis-dependent mRNA stability switch. Their transcripts are stable until mitosis, when a precipitous decay eliminates the mRNA complement, preventing carryover into the next cycle. Remarkably, the specificity and timing of decay is entirely regulated by their promoter, independent of specific cis mRNA sequences. The mitotic exit network protein Dbf2p binds to SWI5 and CLB2 mRNAs cotranscriptionally and regulates their decay. This work reveals the promoter-dependent control of mRNA stability, a regulatory mechanism that could be employed by a variety of mRNAs and organisms.