Heat shock-induced interactions of heat shock transcription factor and the human hsp70 promoter examined by in vivo footprinting.

Genomic footprinting of the human hsp70 promoter reveals that heat shock induces a rapid binding of a factor, presumably heat shock transcription factor, to a region encompassing five contiguous NGAAN sequences, three perfect and two imperfect matches to the consensus sequence. Arrays of inverted NGAAN sequences have been defined as the heat shock element. No protein is bound to the heat shock element prior to or after recovery from heat shock. Heat shock does not perturb the binding of factors to other regulatory elements in the promoter which contribute to basal expression of the hsp70 gene.

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Role of chromatin and Xenopus laevis heat shock transcription factor in regulation of transcription from the X. laevis hsp70 promoter in vivo.

Molecular and Cellular Biology ◽

10.1128/mcb.15.11.6013 ◽

1995 ◽

Vol 15 (11) ◽

pp. 6013-6024 ◽

Cited By ~ 44

Author(s):

N Landsberger ◽

A P Wolffe

Keyword(s):

Transcription Factor ◽

Heat Shock ◽

Xenopus Laevis ◽

Heat Shock Transcription Factor ◽

Chromatin Assembly ◽

Regulation Of Transcription ◽

Wild Type ◽

Hsp70 Promoter ◽

Chromatin Template

Xenopus laevis oocytes activate transcription from the Xenopus hsp70 promoter within a chromatin template in response to heat shock. Expression of exogenous Xenopus heat shock transcription factor 1 (XHSF1) causes the activation of the wild-type hsp70 promoter within chromatin. XHSF1 activates transcription at normal growth temperatures (18 degrees C), but heat shock (34 degrees C) facilitates transcriptional activation. Titration of chromatin in vivo leads to constitutive transcription from the wild-type hsp70 promoter. The Y box elements within the hsp70 promoter facilitate transcription in the presence or absence of chromatin. The presence of the Y box elements prevents the assembly of canonical nucleosomal arrays over the promoter and facilitates transcription. In a mutant hsp70 promoter lacking Y boxes, exogenous XHSF1 activates transcription from a chromatin template much more efficiently under heat shock conditions. Activation of transcription from the mutant promoter by exogenous XHSF1 correlates with the disappearance of a canonical nucleosomal array over the promoter. Chromatin structure on a mutant hsp70 promoter lacking Y boxes can restrict XHSF1 access; however, on both mutant and wild-type promoters, chromatin assembly can also restrict the function of the basal transcriptional machinery. We suggest that chromatin assembly has a physiological role in establishing a transcriptionally repressed state on the Xenopus hsp70 promoter in vivo.

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TATA-dependent and TATA-independent function of the basal and heat shock elements of a human hsp70 promoter

Molecular and Cellular Biology ◽

10.1128/mcb.10.4.1319-1328.1990 ◽

1990 ◽

Vol 10 (4) ◽

pp. 1319-1328

Author(s):

J M Greene ◽

R E Kingston

Keyword(s):

Heat Shock ◽

Consensus Sequence ◽

Simian Virus 40 ◽

Simian Virus ◽

Hsp70 Promoter ◽

Heat Shock Element ◽

Tata Element ◽

Sequence Elements ◽

Human Heat Shock Protein ◽

Human Hsp70

We have characterized the interactions between the TATA element and other sequence elements of a human heat shock protein 70 (hsp70) promoter by a mutational approach. Expression of a distal element of this promoter requires an intact TATA element in human cell lines. The hsp70 TATA element can be functionally replaced for this interaction by TATA elements from the simian virus 40 early and adenovirus EIIa promoters. The TATA element in this promoter therefore both determines the appropriate start site and determines strength by allowing function of the distal element. In contrast, three proximal upstream elements necessary for basal and heat-regulated transcription have no requirement either for a TATA element or for any other proximal element. The behavior of promoters multiply mutant in these proximal elements implies that these elements function independently. We examined the interaction between the heat shock element (HSE) and the TATA element as the distance between the two factor-binding sites was increased. It was necessary to create a mutant HSE with an extended consensus sequence in order for the HSE to function at a distance. Moving this extended HSE 500 bases upstream did not increase its dependence on the TATA element, suggesting that the TATA independence of this element is intrinsic to its function and is not determined by distance from the promoter.

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TATA-dependent and TATA-independent function of the basal and heat shock elements of a human hsp70 promoter.

Molecular and Cellular Biology ◽

10.1128/mcb.10.4.1319 ◽

1990 ◽

Vol 10 (4) ◽

pp. 1319-1328 ◽

Cited By ~ 17

Author(s):

J M Greene ◽

R E Kingston

Keyword(s):

Heat Shock ◽

Consensus Sequence ◽

Simian Virus 40 ◽

Simian Virus ◽

Hsp70 Promoter ◽

Heat Shock Element ◽

Tata Element ◽

Sequence Elements ◽

Human Heat Shock Protein ◽

Human Hsp70

We have characterized the interactions between the TATA element and other sequence elements of a human heat shock protein 70 (hsp70) promoter by a mutational approach. Expression of a distal element of this promoter requires an intact TATA element in human cell lines. The hsp70 TATA element can be functionally replaced for this interaction by TATA elements from the simian virus 40 early and adenovirus EIIa promoters. The TATA element in this promoter therefore both determines the appropriate start site and determines strength by allowing function of the distal element. In contrast, three proximal upstream elements necessary for basal and heat-regulated transcription have no requirement either for a TATA element or for any other proximal element. The behavior of promoters multiply mutant in these proximal elements implies that these elements function independently. We examined the interaction between the heat shock element (HSE) and the TATA element as the distance between the two factor-binding sites was increased. It was necessary to create a mutant HSE with an extended consensus sequence in order for the HSE to function at a distance. Moving this extended HSE 500 bases upstream did not increase its dependence on the TATA element, suggesting that the TATA independence of this element is intrinsic to its function and is not determined by distance from the promoter.

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Cooperative binding of heat shock transcription factor to the Hsp70 promoter in vivo and in vitro.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(17)37615-9 ◽

1994 ◽

Vol 269 (7) ◽

pp. 4804-4811 ◽

Cited By ~ 1

Author(s):

J. Amin ◽

M. Fernandez ◽

J. Ananthan ◽

J.T. Lis ◽

R. Voellmy

Keyword(s):

Transcription Factor ◽

Heat Shock ◽

Heat Shock Transcription Factor ◽

Cooperative Binding ◽

Hsp70 Promoter

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Genome-Wide Analysis of the Biology of Stress Responses through Heat Shock Transcription Factor

Molecular and Cellular Biology ◽

10.1128/mcb.24.12.5249-5256.2004 ◽

2004 ◽

Vol 24 (12) ◽

pp. 5249-5256 ◽

Cited By ~ 297

Author(s):

Ji-Sook Hahn ◽

Zhanzhi Hu ◽

Dennis J. Thiele ◽

Vishwanath R. Iyer

Keyword(s):

Transcription Factor ◽

Heat Shock ◽

Stress Responses ◽

Target Genes ◽

Full Range ◽

Regulatory Elements ◽

Heat Shock Transcription Factor ◽

Metabolic Reprogramming ◽

Heat Shock Element ◽

Genome Wide

ABSTRACT Heat shock transcription factor (HSF) and the promoter heat shock element (HSE) are among the most highly conserved transcriptional regulatory elements in nature. HSF mediates the transcriptional response of eukaryotic cells to heat, infection and inflammation, pharmacological agents, and other stresses. While HSF is essential for cell viability in Saccharomyces cerevisiae, oogenesis and early development in Drosophila melanogaster, extended life span in Caenorhabditis elegans, and extraembryonic development and stress resistance in mammals, little is known about its full range of biological target genes. We used whole-genome analyses to identify virtually all of the direct transcriptional targets of yeast HSF, representing nearly 3% of the genomic loci. The majority of the identified loci are heat-inducibly bound by yeast HSF, and the target genes encode proteins that have a broad range of biological functions including protein folding and degradation, energy generation, protein trafficking, maintenance of cell integrity, small molecule transport, cell signaling, and transcription. This genome-wide identification of HSF target genes provides novel insights into the role of HSF in growth, development, disease, and aging and in the complex metabolic reprogramming that occurs in all cells in response to stress.

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Activation of the DNA-binding ability of human heat shock transcription factor 1 may involve the transition from an intramolecular to an intermolecular triple-stranded coiled-coil structure.

Molecular and Cellular Biology ◽

10.1128/mcb.14.11.7557 ◽

1994 ◽

Vol 14 (11) ◽

pp. 7557-7568 ◽

Cited By ~ 126

Author(s):

J Zuo ◽

R Baler ◽

G Dahl ◽

R Voellmy

Keyword(s):

Transcription Factor ◽

Heat Stress ◽

Heat Shock ◽

Dna Binding ◽

Hydrophobic Interactions ◽

Coiled Coil ◽

Heat Shock Transcription Factor ◽

Single Amino Acid ◽

Binding Ability ◽

Heat Shock Element

Heat stress regulation of human heat shock genes is mediated by human heat shock transcription factor hHSF1, which contains three 4-3 hydrophobic repeats (LZ1 to LZ3). In unstressed human cells (37 degrees C), hHSF1 appears to be in an inactive, monomeric state that may be maintained through intramolecular interactions stabilized by transient interaction with hsp70. Heat stress (39 to 42 degrees C) disrupts these interactions, and hHSF1 homotrimerizes and acquires heat shock element DNA-binding ability. hHSF1 expressed in Xenopus oocytes also assumes a monomeric, non-DNA-binding state and is converted to a trimeric, DNA-binding form upon exposure of the oocytes to heat shock (35 to 37 degrees C in this organism). Because endogenous HSF DNA-binding activity is low and anti-hHSF1 antibody does not recognize Xenopus HSF, we employed this system for mapping regions in hHSF1 that are required for the maintenance of the monomeric state. The results of mutagenesis analyses strongly suggest that the inactive hHSF1 monomer is stabilized by hydrophobic interactions involving all three leucine zippers which may form a triple-stranded coiled coil. Trimerization may enable the DNA-binding function of hHSF1 by facilitating cooperative binding of monomeric DNA-binding domains to the heat shock element motif. This view is supported by observations that several different LexA DNA-binding domain-hHSF1 chimeras bind to a LexA-binding site in a heat-regulated fashion, that single amino acid replacements disrupting the integrity of hydrophobic repeats render these chimeras constitutively trimeric and DNA binding, and that LexA itself binds stably to DNA only as a dimer but not as a monomer in our assays.

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Activation of the heat shock transcription factor by hypoxia in normal and tumor cell lines in vivo and in vitro

International Journal of Radiation Oncology*Biology*Physics ◽

10.1016/0360-3016(92)90667-7 ◽

1992 ◽

Vol 23 (4) ◽

pp. 891-897 ◽

Cited By ~ 19

Author(s):

Amato J. Giaccia ◽

Elizabeth A. Auger ◽

Albert Koong ◽

David J. Terris ◽

Andrew I. Minchinton ◽

...

Keyword(s):

Transcription Factor ◽

Heat Shock ◽

Tumor Cell ◽

Cell Lines ◽

Heat Shock Transcription Factor ◽

Tumor Cell Lines

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Modular recognition of 5-base-pair DNA sequence motifs by human heat shock transcription factor

Molecular and Cellular Biology ◽

10.1128/mcb.11.7.3504-3514.1991 ◽

1991 ◽

Vol 11 (7) ◽

pp. 3504-3514

Author(s):

N F Cunniff ◽

J Wagner ◽

W D Morgan

Keyword(s):

Transcription Factor ◽

Heat Shock ◽

Gene Promoter ◽

Heat Shock Transcription Factor ◽

Heat Shock Gene ◽

Gene Promoters ◽

Sequence Motifs ◽

Binding Modes ◽

Heat Shock Element

We investigated the recognition of the conserved 5-bp repeated motif NGAAN, which occurs in heat shock gene promoters of Drosophila melanogaster and other eukaryotic organisms, by human heat shock transcription factor (HSF). Extended heat shock element mutants of the human HSP70 gene promoter, containing additional NGAAN blocks flanking the original element, showed significantly higher affinity than the wild-type promoter element for human HSF in vitro. Protein-DNA contact positions were identified by hydroxyl radical protection, diethyl pyrocarbonate interference, and DNase I footprinting. New contacts in the mutant HSE constructs corresponded to the locations of additional NGAAN motifs. The pattern of binding indicated the occurrence of multiple DNA binding modes for HSF with the various constructs and was consistent with an oligomeric, possibly trimeric, structure of the protein. In contrast to the improved binding, the extended heat shock element mutant constructs did not exhibit dramatically increased heat-inducible transcription in transient expression assays with HeLa cells.

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Two transcriptional activators, CCAAT-box-binding transcription factor and heat shock transcription factor, interact with a human hsp70 gene promoter

Molecular and Cellular Biology ◽

10.1128/mcb.7.3.1129-1138.1987 ◽

1987 ◽

Vol 7 (3) ◽

pp. 1129-1138

Author(s):

W D Morgan ◽

G T Williams ◽

R I Morimoto ◽

J Greene ◽

R E Kingston ◽

...

Keyword(s):

Transcription Factor ◽

Heat Shock ◽

Gene Promoter ◽

Heat Shock Transcription Factor ◽

Transcriptional Activators ◽

Hsp70 Gene ◽

Nuclear Extracts ◽

Ccaat Box ◽

Human Hsp70

We characterized the activity of a human hsp70 gene promoter by in vitro transcription. Analysis of 5' deletion and substitution mutants in HeLa nuclear extracts showed that the basal activity of the promoter depends primarily on a CCAAT-box sequence located at -65. A protein factor, CCAAT-box-binding transcription factor (CTF), was isolated from HeLa nuclear extracts and shown to be responsible for stimulation of transcription in a reconstituted in vitro system. DNase I footprinting revealed that CTF interacts with two CCAAT-box elements located at -65 and -147 of the human hsp70 promoter. An additional binding activity, heat shock transcription factor (HSTF), which interacted with the heat shock element, was also identified in HeLa extract fractions. This demonstrates that the promoter of this human hsp70 gene interacts with at least two positive transcriptional activators, CTF, which is required for CCAAT-box-dependent transcription as in other promoters such as those of globin and herpes simplex virus thymidine kinase genes, and HSTF, which is involved in heat inducibility.

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