Photofootprinting in vivo detects transcription-dependent changes in yeast TATA boxes

Scott B. Selleck, John Majors

Research output: Contribution to journalArticle

71 Scopus citations

Abstract

To elucidate critical steps in the transcription initiation process, we have devised a protocol for obtaining information about DNA structure and DNA-protein interactions at nucleotide level resolution from intact yeast cells. Our procedure combines the ultraviolet light 'footprinting' method developed by Becker and Wang1 with the 'genomic sequencing' technique described by Church and Gilbert2. Using this approach we were able to detect the binding of GAL 4 protein at sites within the upstream activating sequence (UASG) previously mapped using other in vivo and in vitro foot-printing procedures3-5. We also observed transcription-dependent changes in sensitivity of DNA to ultraviolet-induced covalent modification at several positions between the upstream activating sequence and the transcription initiation sites of the GAL 1 and GAL 10 genes. The most prominent of these changes occurs at a common site within the putative 'TATA' boxes of the two genes. Ultraviolet modification at this site is enhanced only in transcrip-tionally active promoters.

Original languageEnglish (US)
Pages (from-to)173-177
Number of pages5
JournalNature
Volume325
Issue number6100
DOIs
StatePublished - Dec 1 1987

All Science Journal Classification (ASJC) codes

  • General

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