Uncoupling gene activity from chromatin structure: promoter mutations can inactivate transcription of the yeast HSP82 gene without eliminating nucleosome-free regions.
- 1 October 1992
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 89 (19) , 9166-9170
- https://doi.org/10.1073/pnas.89.19.9166
Abstract
DNase I-hypersensitive sites represent "nucleosome-free" regions in chromatin where the underlying DNA sequence is highly accessible to trans-acting proteins. Here we demonstrate that it is possible to uncouple gene activity from hypersensitive site formation. Point or substitution mutations were introduced into the promoter of the yeast chromosomal HSP82 gene, encoding the 83-kDa heat shock protein (HSP), via site-directed integration. Mutating either the TATA box or heat shock element 1 (HSE1) significantly reduced basal and heat-induced transcription while mutating both essentially inactivated expression. Dormant transcription units exhibited arrays of sequence-positioned nucleosomes; nevertheless, the inactivated genes still retained a hypersensitive site within their mutated promoters. In addition, all yeast strains maintained a heat-inducible hypersensitive site at -600 base pairs (bp), while several mutant strains converted a constitutive hypersensitive site at -300 bp into a heat-inducible one. Thus, mutations in cis-acting elements within a promoter can inactivate transcription without eliminating nucleosome-free regions.Keywords
This publication has 48 references indexed in Scilit:
- The complexities of eukaryotic transcription initiation: regulation of preinitiation complex assemblyTrends in Biochemical Sciences, 1991
- A yeast protein that influences the chromatin structure of UASG and functions as a powerful auxiliary gene activator.Genes & Development, 1990
- Complex modes of heat shock factor activation.Molecular and Cellular Biology, 1990
- Basal-level expression of the yeast HSP82 gene requires a heat shock regulatory element.Molecular and Cellular Biology, 1989
- Activation of yeast RNA polymerase II transcription by a thymidine-rich upstream element in vitro.Proceedings of the National Academy of Sciences, 1989
- The chromatin structure at the promoter of a glyceraldehyde phosphate dehydrogenase gene from Saccharomyces cerevisiae reflects its functional state.Molecular and Cellular Biology, 1988
- Constitutive binding of yeast heat shock factor to DNA in vivo.Molecular and Cellular Biology, 1988
- Isolation of the gene encoding the S. cerevisiae heat shock transcription factorCell, 1988
- NUCLEASE HYPERSENSITIVE SITES IN CHROMATINAnnual Review of Biochemistry, 1988
- Heat shock-regulated production of Escherichia coli beta-galactosidase in Saccharomyces cerevisiae.Molecular and Cellular Biology, 1983