Molecular mechanism of attenuation of heat shock transcription factor 1 activity

Transactivation Domain ◽

Proteotoxic Stress ◽

Transcription Factor 1 ◽

Over 40 Years

AbstractThe heat shock response is a universal transcriptional response to proteotoxic stress orchestrated by heat shock transcription factor Hsf1 in all eukaryotic cells. Despite over 40 years of intense research, the mechanism of HSF1 activity regulation remains poorly understood at a molecular level. In metazoa Hsf1 trimerizes upon heat shock through a leucin-zipper domain and binds to DNA. How Hsf1 is dislodged from DNA and monomerized remained enigmatic. Here, we demonstrate that trimeric Hsf1 is dissociated from DNA in vitro by Hsc70 and DnaJB1. Hsc70 acts at two distinct sites on Hsf1. Hsf1 trimers are monomerized by successive cycles of entropic pulling, unzipping the triple leucine-zipper. This process directly monitors the concentration of Hsc70 and DnaJB1. During heat shock adaptation Hsc70 first binds to the transactivation domain leading to partial attenuation of the response and subsequently, at higher concentrations, Hsc70 removes Hsf1 from DNA to restore the resting state.

Heat Shock Element Architecture Is an Important Determinant in the Temperature and Transactivation Domain Requirements for Heat Shock Transcription Factor

10.1128/mcb.18.11.6340 ◽

1998 ◽

Vol 18 (11) ◽

pp. 6340-6352 ◽

Cited By ~ 71

Author(s):

Nicholas Santoro ◽

Nina Johansson ◽

Dennis J. Thiele

Keyword(s):

Transcription Factor ◽

Heat Shock ◽

Transcriptional Activation ◽

Single Gene ◽

Limited Proteolysis ◽

Normal Growth ◽

Transactivation Domain ◽

Heat Shock Element

ABSTRACT The baker’s yeast Saccharomyces cerevisiae possesses a single gene encoding heat shock transcription factor (HSF), which is required for the activation of genes that participate in stress protection as well as normal growth and viability. Yeast HSF (yHSF) contains two distinct transcriptional activation regions located at the amino and carboxyl termini. Activation of the yeast metallothionein gene, CUP1, depends on a nonconsensus heat shock element (HSE), occurs at higher temperatures than other heat shock-responsive genes, and is highly dependent on the carboxyl-terminal transactivation domain (CTA) of yHSF. The results described here show that the noncanonical (or gapped) spacing of GAA units in the CUP1HSE (HSE1) functions to limit the magnitude of CUP1transcriptional activation in response to heat and oxidative stress. The spacing in HSE1 modulates the dependence for transcriptional activation by both stresses on the yHSF CTA. Furthermore, a previously uncharacterized HSE in the CUP1 promoter, HSE2, modulates the magnitude of the transcriptional activation of CUP1, via HSE1, in response to stress. In vitro DNase I footprinting experiments suggest that the occupation of HSE2 by yHSF strongly influences the manner in which yHSF occupies HSE1. Limited proteolysis assays show that HSF adopts a distinct protease-sensitive conformation when bound to the CUP1HSE1, providing evidence that the HSE influences DNA-bound HSF conformation. Together, these results suggest that CUP1regulation is distinct from that of other classic heat shock genes through the interaction of yHSF with two nonconsensus HSEs. Consistent with this view, we have identified other gene targets of yHSF containing HSEs with sequence and spacing features similar to those ofCUP1 HSE1 and show a correlation between the spacing of the GAA units and the relative dependence on the yHSF CTA.

Evaluation of in vitro and in vivo therapeutic antitumor efficacy of transduction of polo‑like kinase 1 and heat shock transcription factor 1 small interfering RNA

Experimental and Therapeutic Medicine ◽

10.3892/etm.2017.5060 ◽

2017 ◽

Cited By ~ 2

Author(s):

Yoshiyuki Hattori ◽

Takuto Kikuchi ◽

Kei‑Ichi Ozaki ◽

Hiraku Onishi

Keyword(s):

Transcription Factor ◽

Heat Shock ◽

Small Interfering Rna ◽

Antitumor Efficacy ◽

Interfering Rna ◽

The Role of Heat Shock Transcription Factor 1 in the Genome-wide Regulation of the Mammalian Heat Shock Response

Molecular Biology of the Cell ◽

10.1091/mbc.e03-10-0738 ◽

2004 ◽

Vol 15 (3) ◽

pp. 1254-1261 ◽

Cited By ~ 222

Author(s):

Nathan D. Trinklein ◽

John I. Murray ◽

Sara J. Hartman ◽

David Botstein ◽

Richard M. Myers

Keyword(s):

Transcription Factor ◽

Heat Shock ◽

Heat Shock Response ◽

Mammalian Cells ◽

Transcriptional Response ◽

Genome Wide ◽

Shock Response ◽

Inducible Genes ◽

Previous work has implicated heat shock transcription factor 1 (HSF1) as the primary transcription factor responsible for the transcriptional response to heat stress in mammalian cells. We characterized the heat shock response of mammalian cells by measuring changes in transcript levels and assaying binding of HSF1 to promoter regions for candidate heat shock genes chosen by a combination of genome-wide computational and experimental methods. We found that many heat-inducible genes have HSF1 binding sites (heat shock elements, HSEs) in their promoters that are bound by HSF1. Surprisingly, for 24 heat-inducible genes, we detected no HSEs and no HSF1 binding. Furthermore, of 182 promoters with likely HSE sequences, we detected HSF1 binding at only 94 of these promoters. Also unexpectedly, we found 48 genes with HSEs in their promoters that are bound by HSF1 but that nevertheless did not show induction after heat shock in the cell types we examined. We also studied the transcriptional response to heat shock in fibroblasts from mice lacking the HSF1 gene. We found 36 genes in these cells that are induced by heat as well as they are in wild-type cells. These results provide evidence that HSF1 does not regulate the induction of every transcript that accumulates after heat shock, and our results suggest that an independent posttranscriptional mechanism regulates the accumulation of a significant number of transcripts.

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 ◽

Tumor Cell Lines

Glycogen Synthase Kinase 3β and Extracellular Signal-Regulated Kinase Inactivate Heat Shock Transcription Factor 1 by Facilitating the Disappearance of Transcriptionally Active Granules after Heat Shock

10.1128/mcb.18.11.6624 ◽

1998 ◽

Vol 18 (11) ◽

pp. 6624-6633 ◽

Cited By ~ 118

Author(s):

Bin He ◽

Yong-Hong Meng ◽

Nahid F. Mivechi

Keyword(s):

Transcription Factor ◽

Heat Shock ◽

Transcriptional Activity ◽

Glycogen Synthase ◽

Glycogen Synthase Kinase 3Β ◽

Extracellular Signal Regulated Kinase ◽

Extracellular Signal ◽

Gsk 3Β ◽

ABSTRACT Heat shock transcription factor 1 (HSF-1) activates the transcription of heat shock genes in eukaryotes. Under normal physiological growth conditions, HSF-1 is a monomer. Its transcriptional activity is repressed by constitutive phosphorylation. Upon activation, HSF-1 forms trimers, acquires DNA binding activity, increases transcriptional activity, and appears as punctate granules in the nucleus. In this study, using bromouridine incorporation and confocal laser microscopy, we demonstrated that newly synthesized pre-mRNAs colocalize to the HSF-1 punctate granules after heat shock, suggesting that these granules are sites of transcription. We further present evidence that glycogen synthase kinase 3β (GSK-3β) and extracellular signal-regulated kinase mitogen-activated protein kinase (ERK MAPK) participate in the down regulation of HSF-1 transcriptional activity. Transient increases in the expression of GSK-3β facilitate the disappearance of HSF-1 punctate granules and reduce hsp-70 transcription after heat shock. We have also shown that ERK is the priming kinase for GSK-3β. Taken together, these results indicate that GSK-3β and ERK MAPK facilitate the inactivation of activated HSF-1 after heat shock by dispersing HSF-1 from the sites of transcription.

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

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 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.

Homology Modeling and Characterization of Novel Stress Inducing Genes and their Split Sites Recognition in the Genome of Zebra Fish (Danio Rario) by Employing Special Consensus Pattern Matching Algorithms- using an in-Silico Method.

International Journal of Engineering and Advanced Technology - Regular Issue ◽

10.35940/ijeat.a2003.109119 ◽

2019 ◽

Vol 9 (1) ◽

pp. 6715-6720

Keyword(s):

Transcription Factor ◽

Heat Shock ◽

Pattern Matching ◽

Binding Sites ◽

In Silico ◽

3D Structure ◽

Transcription Factor Binding Sites ◽

Zebra Fish ◽

Zebra fish has long been considered to be as a strong animal model in biology and modern genetics; however now a days its gaining lot of importance in environmental studies as well. The readily availability of entire genome sequences made to permit carrying out in silico studies at Genomic level. As everyone is known that stress is much more complex and complicated process that involves so much of gene regulations known as up regulation and down regulation, the corresponding stress proteins, broadly known as heat shock proteins. In the current study, the potential transcription factor binding sites were traced out by using bioinformatics tools and about 50 heat shock protein genes were predicted by using special alogorithms using pattern matching and position weight matrices. The 3D structure of DNA-binding domain of HSTF-1 ( Heat Shock Transcription factor-1) which is crucial for regulating heat shot proteins was traced out and builted by using homology modelling methods. The 3D structure of the heat shock transcription factor-1 and together with predicted transcription factor binding sites may be validated in future experimental works which would help us in understanding the complex responsive stress mechanisms lying in Zebra fish.

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

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 ◽

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.