Home LiteratureArticle Details
PMID: 1409619 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Uncoupling gene activity from chromatin structure: promoter mutations can inactivate transcription of the yeast HSP82 gene without eliminating nucleosome-free regions.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 89 ·No. 19 ·1992-10-01 ·Pages 9166-70

Lee MS, Garrard WT

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.

Related Genes
HSP82
MeSH Terms
Alleles Base Sequence Binding Sites DNA, Fungal/genetics,metabolism Deoxyribonuclease I Gene Expression Regulation, Fungal Genes, Fungal Heat-Shock Proteins/genetics Hot Temperature Kinetics Molecular Sequence Data Mutagenesis, Site-Directed Oligodeoxyribonucleotides Promoter Regions, Genetic RNA, Messenger/metabolism Saccharomyces cerevisiae/genetics TATA Box Transcription, Genetic
Chemicals
DNA, Fungal Heat-Shock Proteins Oligodeoxyribonucleotides RNA, Messenger Deoxyribonuclease I
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Lee M S
Department of Microbiology, University of Ulsan, Kyongnam, Korea.
Garrard W T
References (43)
43 references, click to expand
  1. The complexities of eukaryotic transcription initiation: regulation of preinitiation complex assembly.
    Trends Biochem Sci. 1991 Nov;16(11):402-8 PMID: 1776168
  2. Heat shock-regulated production of Escherichia coli beta-galactosidase in Saccharomyces cerevisiae.
    Mol Cell Biol. 1983 Sep;3(9):1625-33 PMID: 6415404
  3. A yeast protein that influences the chromatin structure of UASG and functions as a powerful auxiliary gene activator.
    Genes Dev. 1990 Apr;4(4):503-14 PMID: 2361590
  4. Complex modes of heat shock factor activation.
    Mol Cell Biol. 1990 Feb;10(2):752-9 PMID: 2405254
  5. Activation of yeast RNA polymerase II transcription by a thymidine-rich upstream element in vitro.
    Proc Natl Acad Sci U S A. 1989 Jan;86(2):486-90 PMID: 2643115
  6. A DNA binding protein that recognizes oligo(dA).oligo(dT) tracts.
    EMBO J. 1989 Jun;8(6):1867-77 PMID: 2670564
  7. Basal-level expression of the yeast HSP82 gene requires a heat shock regulatory element.
    Mol Cell Biol. 1989 Nov;9(11):4789-98 PMID: 2689867
  8. The chromatin structure at the promoter of a glyceraldehyde phosphate dehydrogenase gene from Saccharomyces cerevisiae reflects its functional state.
    Mol Cell Biol. 1988 Dec;8(12):5513-20 PMID: 2854200
  9. Isolation of the gene encoding the S. cerevisiae heat shock transcription factor.
    Cell. 1988 Sep 9;54(6):841-53 PMID: 3044612
  10. Nuclease hypersensitive sites in chromatin.
    Annu Rev Biochem. 1988;57:159-97 PMID: 3052270
  11. Constitutive binding of yeast heat shock factor to DNA in vivo.
    Mol Cell Biol. 1988 Nov;8(11):5040-2 PMID: 3062378
  12. Germline transformation used to define key features of heat-shock response elements.
    Science. 1988 Mar 4;239(4844):1139-42 PMID: 3125608
  13. Two DNA-binding factors recognize specific sequences at silencers, upstream activating sequences, autonomously replicating sequences, and telomeres in Saccharomyces cerevisiae.
    Mol Cell Biol. 1988 Jan;8(1):210-25 PMID: 3275867
  14. Sharp boundaries demarcate the chromatin structure of a yeast heat-shock gene.
    J Mol Biol. 1987 Jan 5;193(1):71-80 PMID: 3295258
  15. Heat shock factor is regulated differently in yeast and HeLa cells.
    Nature. 1987 Sep 3-9;329(6134):81-4 PMID: 3306402
  16. Purification and cloning of a DNA binding protein from yeast that binds to both silencer and activator elements.
    Cell. 1987 Dec 4;51(5):721-32 PMID: 3315231
  17. Protein-DNA interactions and nuclease-sensitive regions determine nucleosome positions on yeast plasmid chromatin.
    J Mol Biol. 1986 Jul 20;190(2):177-90 PMID: 3540310
  18. The relationship of regulatory proteins and DNase I hypersensitive sites in the yeast GAL1-10 genes.
    Nucleic Acids Res. 1985 Dec 9;13(23):8409-23 PMID: 3909104
  19. Regulation of genes controlling synthesis of the galactose pathway enzymes in yeast.
    Genetics. 1966 Sep;54(3):911-6 PMID: 5970626
  20. The 5' ends of Drosophila heat shock genes in chromatin are hypersensitive to DNase I.
    Nature. 1980 Aug 28;286(5776):854-60 PMID: 6774262
  21. Two types of TATA elements for the CYC1 gene of the yeast Saccharomyces cerevisiae.
    Mol Cell Biol. 1991 Feb;11(2):666-76 PMID: 1846668
  22. Transcription-induced nucleosome 'splitting': an underlying structure for DNase I sensitive chromatin.
    EMBO J. 1991 Mar;10(3):607-15 PMID: 2001676
  23. Transcriptional regulation of a yeast HSP70 gene by heat shock factor and an upstream repression site-binding factor.
    Genes Dev. 1991 Jul;5(7):1299-308 PMID: 2065978
  24. Role of trans-activating proteins in the generation of active chromatin at the PHO5 promoter in S. cerevisiae.
    EMBO J. 1990 Aug;9(8):2523-8 PMID: 2196175
  25. Yeast heat shock factor contains separable transient and sustained response transcriptional activators.
    Cell. 1990 Aug 24;62(4):793-805 PMID: 2201452
  26. TFIID can be rate limiting in vivo for TATA-containing, but not TATA-lacking, RNA polymerase II promoters.
    Genes Dev. 1992 Feb;6(2):304-15 PMID: 1737620
  27. Positive DNA supercoiling generates a chromatin conformation characteristic of highly active genes.
    Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9675-9 PMID: 1946386
  28. Heat shock factor and the heat shock response.
    Cell. 1991 May 3;65(3):363-6 PMID: 2018972
  29. Genomic footprinting of the yeast HSP82 promoter reveals marked distortion of the DNA helix and constitutive occupancy of heat shock and TATA elements.
    J Mol Biol. 1990 Dec 5;216(3):611-31 PMID: 2175361
  30. The yeast heat shock transcription factor contains a transcriptional activation domain whose activity is repressed under nonshock conditions.
    Cell. 1990 Aug 24;62(4):807-17 PMID: 2201453
  31. Position effect at S. cerevisiae telomeres: reversible repression of Pol II transcription.
    Cell. 1990 Nov 16;63(4):751-62 PMID: 2225075
  32. Yeast TATA-binding protein TFIID binds to TATA elements with both consensus and nonconsensus DNA sequences.
    Proc Natl Acad Sci U S A. 1989 Aug;86(15):5718-22 PMID: 2569738
  33. Positive and negative regulation of basal expression of a yeast HSP70 gene.
    Mol Cell Biol. 1989 May;9(5):2025-33 PMID: 2664467
  34. hsp82 is an essential protein that is required in higher concentrations for growth of cells at higher temperatures.
    Mol Cell Biol. 1989 Sep;9(9):3919-30 PMID: 2674684
  35. Protein/DNA architecture of the DNase I hypersensitive region of the Drosophila hsp26 promoter.
    EMBO J. 1988 Jul;7(7):2191-201 PMID: 2901349
  36. Yeast heat shock factor is an essential DNA-binding protein that exhibits temperature-dependent phosphorylation.
    Cell. 1988 Sep 9;54(6):855-64 PMID: 3044613
  37. Statistical positioning of nucleosomes by specific protein-binding to an upstream activating sequence in yeast.
    J Mol Biol. 1988 Nov 5;204(1):109-27 PMID: 3063825
  38. Key features of heat shock regulatory elements.
    Mol Cell Biol. 1988 Sep;8(9):3761-9 PMID: 3146692
  39. Saturation mutagenesis of a yeast his3 "TATA element": genetic evidence for a specific TATA-binding protein.
    Proc Natl Acad Sci U S A. 1988 Apr;85(8):2691-5 PMID: 3282236
  40. In vivo DNA-binding properties of a yeast transcription activator protein.
    Mol Cell Biol. 1987 Sep;7(9):3260-7 PMID: 3313011
  41. Purification and characterization of a heat-shock element binding protein from yeast.
    EMBO J. 1987 Oct;6(10):3035-41 PMID: 3319580
  42. Naturally occurring poly(dA-dT) sequences are upstream promoter elements for constitutive transcription in yeast.
    Proc Natl Acad Sci U S A. 1985 Dec;82(24):8419-23 PMID: 3909145
  43. Regulatory DNA-binding proteins in yeast: an overview.
    Yeast. 1990 Jul-Aug;6(4):271-97 PMID: 2204245
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.89.19.9166
Published
1992-10-01
Pages
9166-70
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC50086
Subset
IM
Grants
NIGMS NIH HHS · GM22201 · United States
NIGMS NIH HHS · GM29935 · United States
NIGMS NIH HHS · GM31689 · United States
External Links
PubMed source DOI: 10.1073/pnas.89.19.9166 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2