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.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Becker, M. M. & Wang, J. C. Nature 309, 682–687 (1984).
Church, G. M. & Gilbert, W. Proc. nat. Acad. Sci. U.S.A. 81, 1991–1995 (1984).
Giniger, E., Varnum, S. M. & Ptashne, M. Cell 40, 767–774 (1985).
Bram, R. J. & Kornberg, R. D. Proc. nat. Acad. Sci. U.S.A. 82, 43–47 (1985).
Keegan, L., Gill, G. & Ptashne, M. Science 231, 699–704 (1986).
Douglas, H. C. & Hawthorne, D. Genetics 49, 837–844 (1964).
Oshima, Y. The Molecular Biology of the Yeast Saccharomyces: Metabolism and Gene Expression (eds Strathern, J. N., Jones, E. W. & Broach, J. R.) 159–180 (Cold Spring Harbor Press, New York, 1982).
Douglas, H. C. & Pelroy, G. Biochim. biophys. Acta 68, 155–156 (1963).
Johnston, M. & Dover, J. Proc. natn. Acad. Sci. U.S.A. (in the press).
Struhl, K. Proc. natn. Acad. Sci. U.S.A. 79, 7385–7389 (1982).
Guarente, L., Yocum, R. R. & Gifford, P. Proc. natn. Acad. Sci. U.S.A. 79, 7410–7414 (1982).
Guarente, L. & Mason, T. Cell 32, 1279–1286 (1983).
Nagawa, F. & Fink, G. R. Proc. natn. Acad. Sci. U.S.A. 82, 8557–8561 (1985).
Hahn, S., Hoar, E. T. & Guarente, L. Proc. natn. Acad. Sci. U.S.A. 82, 8562–8566 (1985).
McNeil, J. B. & Smith, M. J. molec. Biol. 187, 363–378 (1986).
Maxam, A. M. & Gilbert, W. Proc. natn. Acad. Sci. U.S.A. 74, 560–564 (1977).
Johnston, M. & Davis, R. W. Molec. cell Biol. 4, 1440–1448 (1984).
Carlson, M. & Botstein, D. Cell 28, 145–154 (1982).
Sarokin, L. & Carlson, M. Molec. cell. Biol. 4, 2750–2757 (1984).
West, R. W., Yocum, R. R. & Ptashne, M. Molec. cell. Biol. 4, 2467–2478 (1984).
Breathnach, R. & Chambon, P. A. Rev. Biochem. 50, 349–383 (1981).
Parker, C. S. & Topol, J. Cell 36, 357–369 (1984).
Sawadogo, M. & Roeder, R. G. Cell 43, 165–175 (1985).
Coen, D. M., Weinheimer, S. P. & Mcknight, S. L. Science 234, 53–59 (1986).
Ptashne, M. Nature 322, 697–701 (1986).
Yocum, R. R. & Johnston, M. Gene 32, 75–82 (1984).
Scott, J. H. & Schekman, R. J. Bact. 142, 414–423 (1980).
Tabor, S. & Richardson, C. C. Proc. natn. Acad. Sci. U.S.A. 82, 1074–1078 (1985).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Selleck, S., Majors, J. Photofootprinting in vivo detects transcription-dependent changes in yeast TATA boxes. Nature 325, 173–177 (1987). https://doi.org/10.1038/325173a0
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/325173a0
This article is cited by
-
Base-resolution UV footprinting by sequencing reveals distinctive damage signatures for DNA-binding proteins
Nature Communications (2023)
-
Binding of TBP to promoters in vivo is stimulated by activators and requires Pol II holoenzyme
Nature (1999)
-
Chromatin structure of the yeast SUC2 promoter in regulatory mutants
Molecular and General Genetics MGG (1992)
-
Functional relationship among TATA sequences, gene induction and transcription initiation in the ?-galactosidase, LAC4, gene from Kluyveromyces lactis
Current Genetics (1989)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.