Epitope-tagged yeast strains reveal promoter driven changes to 3'-end formation and convergent antisense-transcription from common 3' UTRs

Angavai Swaminathan, Traude H Beilharz

Research output: Contribution to journalArticleResearchpeer-review

Abstract

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.
Original languageEnglish
Pages (from-to)377-386
Number of pages10
JournalNucleic Acids Research
Volume44
Issue number1
DOIs
Publication statusPublished - 8 Jan 2016

Cite this

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title = "Epitope-tagged yeast strains reveal promoter driven changes to 3'-end formation and convergent antisense-transcription from common 3' UTRs",
abstract = "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.",
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Epitope-tagged yeast strains reveal promoter driven changes to 3'-end formation and convergent antisense-transcription from common 3' UTRs. / Swaminathan, Angavai; Beilharz, Traude H.

In: Nucleic Acids Research, Vol. 44, No. 1, 08.01.2016, p. 377-386.

Research output: Contribution to journalArticleResearchpeer-review

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