Epitope-tagged yeast strains reveal promoter driven changes to 3'-end formation and convergent antisense-transcription from common 3' UTRs.
Epitope-tagging by homologous recombination is ubiquitously used to study gene expression, protein localization and function in yeast. This is generally thought to insulate the regulation of gene expression to that mediated by the promoter and coding regions because native 3' UTR are replaced. Here we show that the 3' UTRs, CYC1 and ADH1, contain cryptic promoters that generate abundant convergent antisense-transcription in Saccharomyces cerevisiae. Moreover we show that aberrant, truncating 3' -end formation is often associated with regulated transcription in TAP-tagged strains. Importantly, the steady-state level of both 3' -truncated and antisense transcription products is locus dependent. Using TAP and GFP-tagged strains we show that the transcriptional state of the gene-of-interest induces changes to 3' -end formation by alternative polyadenylation and antisense transcription from a universal 3' UTR. This means that these 3' UTRs contains plastic features that can be molded to reflect the regulatory architecture of the locus rather than bringing their own regulatory paradigm to the gene-fusions as would be expected. Our work holds a cautionary note for studies utilizing tagged strains for quantitative biology, but also provides a new model for the study of promoter driven rewiring of 3' -end formation and regulatory non-coding transcription.
Development and stem cells Program, Monash Biomedicine Discovery Institute and Department of Biochemistry and Molecular Biology, Monash University, Melbourne, Victoria 3800, Australia.
Angavai
A
Swaminathan
Development and stem cells Program, Monash Biomedicine Discovery Institute and Department of Biochemistry and Molecular Biology, Monash University, Melbourne, Victoria 3800, Australia traude.beilharz@monash.edu.
Traude H
http://orcid.org/0000-0002-8942-9502
TH
Beilharz
D020413:3' Untranslated Regions; D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000378:metabolism; D045304:Cytochromes c / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D056426:Genetic Loci; D026723:Polyadenylation; D011401:Promoter Regions, Genetic; D016372:RNA, Antisense; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D015003:Yeasts / Q000235:genetics
D020413:3' Untranslated Regions; C014059:CYC1 protein, S cerevisiae; D016372:RNA, Antisense; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D045304:Cytochromes c; C516801:ADH1 protein, S cerevisiae; D000426:Alcohol Dehydrogenase