Expression of a constitutively active mutant of heat shock factor 1 under the control of testis-specific hst70 gene promoter in transgenic mice induces degeneration of seminiferous epithelium.

Heat shock activates in somatic cells a set of genes encoding heat shock proteins which function as molecular chaperones. The basic mechanism by which these genes are activated is the interaction of the specific transcription factor HSF1 with a regulatory DNA sequence called heat shock element (HSE). In higher eukaryotes HSF1 is present in unstressed cells as inactive monomers which, in response to cellular stress, aggregate into transcriptionally competent homotrimers. In the present paper we showed that the expression of a transgene encoding mutated constitutively active HSF1 placed under the control of a spermatocyte-specific promoter derived from the hst70 gene severely affects spermatogenesis. We found the testes of transgenic mice to be significantly smaller than those of wild-type males and histological analysis showed massive degeneration of the seminiferous epithelium. The lumen of tubules was devoid of spermatids and spermatozoa and using the TUNEL method we demonstrated a high rate of spermatocyte apoptosis. The molecular mechanism by which constitutively active HSF1 arrests spermatogenesis is not known so far. One can assume that HSF1 can either induce or repress so far unknown target genes involved in germ cell apoptosis.

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Up-regulation of the clusterin gene after proteotoxic stress: implication of HSF1–HSF2 heterocomplexes

Biochemical Journal ◽

10.1042/bj20051190 ◽

2006 ◽

Vol 395 (1) ◽

pp. 223-231 ◽

Cited By ~ 86

Author(s):

Fabien Loison ◽

Laure Debure ◽

Philippe Nizard ◽

Pascale le Goff ◽

Denis Michel ◽

...

Keyword(s):

Heat Shock ◽

Neurodegenerative Diseases ◽

Gel Filtration ◽

Gene Promoter ◽

Proteasome Inhibition ◽

Protein Accumulation ◽

Heat Shock Factor 1 ◽

Mrna Levels ◽

Mobility Shift ◽

Heat Shock Element

Clusterin is a secreted protein chaperone up-regulated in several pathologies, including cancer and neurodegenerative diseases. The present study shows that accumulation of aberrant proteins, caused by the proteasome inhibitor MG132 or the incorporation of the amino acid analogue AZC (L-azetidine-2-carboxylic acid), increased both clusterin protein and mRNA levels in the human glial cell line U-251 MG. Consistently, MG132 treatment was capable of stimulating a 1.3 kb clusterin gene promoter. Promoter deletion and mutation studies revealed a critical MG132-responsive region between −218 and −106 bp, which contains a particular heat-shock element, named CLE for ‘clusterin element’. Gel mobility-shift assays demonstrated that MG132 and AZC treatments induced the formation of a protein complex that bound to CLE. As shown by supershift and chromatin-immunoprecipitation experiments, CLE is bound by HSF1 (heat-shock factor 1) and HSF2 upon proteasome inhibition. Furthermore, co-immunoprecipitation assays indicated that these two transcription factors interact. Gel-filtration analyses revealed that the HSF1–HSF2 heterocomplexes bound to CLE after proteasome inhibition have the same apparent mass as HSF1 homotrimers after heat shock, suggesting that HSF1 and HSF2 could heterotrimerize. Therefore these studies indicate that the clusterin is a good candidate to be part of a cellular defence mechanism against neurodegenerative diseases associated with misfolded protein accumulation or decrease in proteasome activity.

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Corrigendum to: Heat shock factor 1 upregulates transcription of Epstein–Barr Virus nuclear antigen 1 by binding to a heat shock element within the BamHI-Q promoter [Virology 421 (2011) 184–191]

Virology ◽

10.1016/j.virol.2021.06.003 ◽

2021 ◽

Author(s):

Feng-Wei Wang ◽

Xian-Rui Wu ◽

Wen-Ju Liu ◽

Yi-Ji Liao ◽

Sheng Lin ◽

...

Keyword(s):

Heat Shock ◽

Epstein Barr Virus ◽

Heat Shock Factor ◽

Nuclear Antigen ◽

Heat Shock Factor 1 ◽

Heat Shock Element ◽

Barr Virus ◽

Epstein Barr

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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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Regulation of human heme oxygenase-1 gene expression under thermal stress

Blood ◽

10.1182/blood.v87.12.5074.bloodjournal87125074 ◽

1996 ◽

Vol 87 (12) ◽

pp. 5074-5084 ◽

Cited By ~ 99

Author(s):

S Okinaga ◽

K Takahashi ◽

K Takeda ◽

M Yoshizawa ◽

H Fujita ◽

...

Keyword(s):

Gene Expression ◽

Thermal Stress ◽

Heat Shock ◽

Transient Expression ◽

Reporter Gene ◽

Heme Oxygenase ◽

Gene Promoter ◽

Heme Oxygenase 1 ◽

Heat Shock Element ◽

Hsp70 Mrna

Heme oxygenase-1 is an essential enzyme in heme catabolism, and its human gene promoter contains a putative heat shock element (HHO-HSE). This study was designed to analyze the regulation of human heme oxygenase-1 gene expression under thermal stress. The amounts of heme oxygenase-1 protein were not increased by heat shock (incubation at 42 degrees C) in human alveolar macrophages and in a human erythroblastic cell line, YN-1–0-A, whereas heat shock protein 70 (HSP70) was noticeably induced. However, heat shock factor does bind in vitro to HHO-HSE and the synthetic HHO-HSE by itself is sufficient to confer the increase in the transient expression of a reporter gene upon heat shock. The deletion of the sequence, located downstream from HHO-HSE, resulted in the activation of a reporter gene by heat shock. These results suggest that HHO-HSE is potentially functional but is repressed in vivo. Interestingly, heat shock abolished the remarkable increase in the levels of heme oxygenase-1 mRNA in YN-1–0-A cells treated with hemin or cadmium, in which HSP70 mRNA was noticeably induced. Furthermore, transient expression assays showed that heat shock inhibits the cadmium-mediated activation of the heme oxygenase-1 promoter, whereas the HSP70 gene promoter was activated upon heat shock. Such regulation of heme oxygenase-1 under thermal stress may be of physiologic significance in erythroid cells.

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HSF1Base: A Comprehensive Database of HSF1 (Heat Shock Factor 1) Target Genes

International Journal of Molecular Sciences ◽

10.3390/ijms20225815 ◽

2019 ◽

Vol 20 (22) ◽

pp. 5815 ◽

Cited By ~ 5

Author(s):

Kovács ◽

Sigmond ◽

Hotzi ◽

Bohár ◽

Fazekas ◽

...

Keyword(s):

Heat Shock ◽

High Throughput ◽

Ribosome Biogenesis ◽

Target Genes ◽

Enrichment Analysis ◽

Cell Types ◽

Heat Shock Factor ◽

Heat Shock Factor 1 ◽

Genes Encoding ◽

Shock Proteins

: HSF1 (heat shock factor 1) is an evolutionarily conserved master transcriptional regulator of the heat shock response (HSR) in eukaryotic cells. In response to high temperatures, HSF1 upregulates genes encoding molecular chaperones, also called heat shock proteins, which assist the refolding or degradation of damaged intracellular proteins. Accumulating evidence reveals however that HSF1 participates in several other physiological and pathological processes such as differentiation, immune response, and multidrug resistance, as well as in ageing, neurodegenerative demise, and cancer. To address how HSF1 controls these processes one should systematically analyze its target genes. Here we present a novel database called HSF1Base (hsf1base.org) that contains a nearly comprehensive list of HSF1 target genes identified so far. The list was obtained by manually curating publications on individual HSF1 targets and analyzing relevant high throughput transcriptomic and chromatin immunoprecipitation data derived from the literature and the Yeastract database. To support the biological relevance of HSF1 targets identified by high throughput methods, we performed an enrichment analysis of (potential) HSF1 targets across different tissues/cell types and organisms. We found that general HSF1 functions (targets are expressed in all tissues/cell types) are mostly related to cellular proteostasis. Furthermore, HSF1 targets that are conserved across various animal taxa operate mostly in cellular stress pathways (e.g., autophagy), chromatin remodeling, ribosome biogenesis, and ageing. Together, these data highlight diverse roles for HSF1, expanding far beyond the HSR.

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Constitutively active heat shock factor 1 enhances glucose-driven insulin secretion

Metabolism ◽

10.1016/j.metabol.2010.07.029 ◽

2011 ◽

Vol 60 (6) ◽

pp. 789-798 ◽

Cited By ~ 5

Author(s):

Tsuyoshi Uchiyama ◽

Shoichi Tomono ◽

Toshihiro Utsugi ◽

Yoshio Ohyama ◽

Tetsuya Nakamura ◽

...

Keyword(s):

Insulin Secretion ◽

Heat Shock ◽

Heat Shock Factor ◽

Heat Shock Factor 1 ◽

Constitutively Active

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Genetic and epigenetic determinants establish a continuum of Hsf1 occupancy and activity across the yeast genome

Molecular Biology of the Cell ◽

10.1091/mbc.e18-06-0353 ◽

2018 ◽

Vol 29 (26) ◽

pp. 3168-3182 ◽

Cited By ~ 17

Author(s):

David Pincus ◽

Jayamani Anandhakumar ◽

Prathapan Thiru ◽

Michael J. Guertin ◽

Alexander M. Erkine ◽

...

Keyword(s):

Heat Shock ◽

Chromatin Immunoprecipitation ◽

Fold Increase ◽

Yeast Genome ◽

Heat Shock Factor 1 ◽

Rna Seq ◽

Heat Shock Element ◽

Cis Acting ◽

Chromatin Immunoprecipitation Sequencing ◽

Pioneer Factors

Heat shock factor 1 is the master transcriptional regulator of molecular chaperones and binds to the same cis-acting heat shock element (HSE) across the eukaryotic lineage. In budding yeast, Hsf1 drives the transcription of ∼20 genes essential to maintain proteostasis under basal conditions, yet its specific targets and extent of inducible binding during heat shock remain unclear. Here we combine Hsf1 chromatin immunoprecipitation sequencing (seq), nascent RNA-seq, and Hsf1 nuclear depletion to quantify Hsf1 binding and transcription across the yeast genome. We find that Hsf1 binds 74 loci during acute heat shock, and these are linked to 46 genes with strong Hsf1-dependent expression. Notably, Hsf1’s induced DNA binding leads to a disproportionate (∼7.5-fold) increase in nascent transcription. Promoters with high basal Hsf1 occupancy have nucleosome-depleted regions due to the presence of “pioneer factors.” These accessible sites are likely critical for Hsf1 occupancy as the activator is incapable of binding HSEs within a stably positioned, reconstituted nucleosome. In response to heat shock, however, Hsf1 accesses nucleosomal sites and promotes chromatin disassembly in concert with the Remodels Structure of Chromatin (RSC) complex. Our data suggest that the interplay between nucleosome positioning, HSE strength, and active Hsf1 levels allows cells to precisely tune expression of the proteostasis network.

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