title: Single-molecule mRNA decay measurements reveal promoter- regulated mRNA stability in yeast.
abstract: 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.
authors.affiliation: Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors.forename: Tatjana
authors.initials: T
authors.lastname: Trcek
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Larson
authors.forename: Alberto
authors.initials: A
authors.lastname: Moldón
authors.forename: Charles C
authors.initials: CC
authors.lastname: Query
authors.forename: Robert H
authors.initials: RH
authors.lastname: Singer
mesh_terms: D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D019926:Cyclin B / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D017404:In Situ Hybridization, Fluorescence; D007700:Kinetics; D011401:Promoter Regions, Genetic; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D020871:RNA Stability; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics; D014158:Transcription, Genetic
chemical_list: C093749:CLB2 protein, S cerevisiae; D018797:Cell Cycle Proteins; D019926:Cyclin B; C080218:SWI5 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C085768:DBF2 protein, S cerevisiae; C085769:DBF20 protein, S cerevisiae; D017346:Protein Serine-Threonine Kinases