Inducers of the heat shock response stimulate phospholipase C and phospholipase A2 activity in mammalian cells

ABSTRACT So far attenuation of pathogens has been mainly obtained by chemical or heat treatment of microbial pathogens. Recently, live attenuated strains have been produced by genetic modification. We have previously demonstrated that in several prokaryotes as well as in yeasts and mammalian cells the heat shock response is controlled by the membrane physical state (MPS). We have also shown that in Salmonella enterica serovar Typhimurium LT2 (Salmonella Typhimurium) overexpression of a Δ12-desaturase gene alters the MPS, inducing a sharp impairment of transcription of major heat shock genes and failure of the pathogen to grow inside macrophage (MΦ) (A. Porta et al., J. Bacteriol. 192:1988-1998, 2010). Here, we show that overexpression of a homologous Δ9-desaturase sequence in the highly virulent G217B strain of the human fungal pathogen Histoplasma capsulatum causes loss of its ability to survive and persist within murine MΦ along with the impairment of the heat shock response. When the attenuated strain of H. capsulatum was injected in a mouse model of infection, it did not cause disease. Further, treated mice were protected when challenged with the virulent fungal parental strain. Attenuation of virulence in MΦ of two evolutionarily distant pathogens was obtained by genetic modification of the MPS, suggesting that this is a new method that may be used to produce attenuation or loss of virulence in both other intracellular prokaryotic and eukaryotic pathogens. This new procedure to generate attenuated forms of pathogens may be used eventually to produce a novel class of vaccines based on the genetic manipulation of a pathogen's membrane fluid state and stress response.

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Disruption of the three cytoskeletal networks in mammalian cells does not affect transcription, translation, or protein translocation changes induced by heat shock.

Molecular and Cellular Biology ◽

10.1128/mcb.5.7.1571 ◽

1985 ◽

Vol 5 (7) ◽

pp. 1571-1581 ◽

Cited By ~ 56

Author(s):

W J Welch ◽

J R Feramisco

Keyword(s):

Heat Shock ◽

Heat Shock Response ◽

Mammalian Cells ◽

Stress Proteins ◽

Protein Translocation ◽

Shock Response ◽

Cytochalasin E ◽

Cytoskeletal Networks ◽

Shock Proteins ◽

Cytoskeletal Elements

Mammalian cells show a complex series of transcriptional and translational switching events in response to heat shock treatment which ultimately lead to the production and accumulation of a small number of proteins, the so-called heat shock (or stress) proteins. We investigated the heat shock response in both qualitative and quantitative ways in cells that were pretreated with drugs that specifically disrupt one or more of the three major cytoskeletal networks. (These drugs alone, cytochalasin E and colcemid, do not result in induction of the heat shock response.) Our results indicated that disruption of the actin microfilaments, the vimentin-containing intermediate filaments, or the microtubules in living cells does not hinder the ability of the cell to undergo an apparently normal heat shock response. Even when all three networks were simultaneously disrupted (resulting in a loose, baglike appearance of the cells), the cells still underwent a complete heat shock response as assayed by the appearance of the heat shock proteins. In addition, the major induced 72-kilodalton heat shock protein was efficiently translocated from the cytoplasm into its proper location in the nucleus and nucleolus irrespective of the condition of the three cytoskeletal elements.

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Poly(ADP-ribosyl)ation regulates heat shock factor-1 activity and the heat shock response in murine fibroblasts

Biochemistry and Cell Biology ◽

10.1139/o06-083 ◽

2006 ◽

Vol 84 (5) ◽

pp. 703-712 ◽

Cited By ~ 14

Author(s):

Silvia Fossati ◽

Laura Formentini ◽

Zhao-Qi Wang ◽

Flavio Moroni ◽

Alberto Chiarugi

Keyword(s):

Heat Shock ◽

Heat Shock Response ◽

Mammalian Cells ◽

Molecular Mechanisms ◽

Heat Shock Factor ◽

Binding Motif ◽

Heat Shock Factor 1 ◽

Shock Response ◽

Parp 1

Poly(ADP-ribose) polymerase-1 (PARP-1)-dependent poly(ADP-ribose) formation is emerging as a key regulator of transcriptional regulation, even though the targets and underlying molecular mechanisms have not yet been clearly identified. In this study, we gathered information on the role of PARP-1 activity in the heat shock response of mouse fibroblasts. We show that DNA binding of heat shock factor (HSF)-1 was impaired by PARP-1 activity in cellular extracts, and was higher in PARP-1−/− than in PARP-1+/+ cells. No evidence for HSF-1 poly(ADP-ribosyl)ation or PARP-1 interaction was found, but a poly(ADP-ribose) binding motif was identified in the transcription factor amino acid sequence. Consistent with data on HSF-1, the expression of heat-shock protein (HSP)-70 and HSP–27 was facilitated in cells lacking PARP-1. Thermosensitivity, however, was higher in PARP-1−/− than in PARP-1+/+ cells. Accordingly, we report that heat-shocked PARP-1 null fibroblasts showed an increased activation of proapoptotic JNK and decreased transcriptional efficiency of prosurvival NF-κB compared with wild-type counterparts. The data indicate that poly(ADP-ribosyl)ation finely regulates HSF-1 activity, and emphasize the complex role of PARP-1 in the heat-shock response of mammalian cells.

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The translation elongation factor eEF1A1 couples transcription to translation during heat shock response

eLife ◽

10.7554/elife.03164 ◽

2014 ◽

Vol 3 ◽

Cited By ~ 69

Author(s):

Maria Vera ◽

Bibhusita Pani ◽

Lowri A Griffiths ◽

Christian Muchardt ◽

Catherine M Abbott ◽

...

Keyword(s):

Heat Shock ◽

Rna Polymerase Ii ◽

Heat Shock Response ◽

Mammalian Cells ◽

Elongation Factor ◽

Translation Elongation Factor ◽

Translation Elongation ◽

Hsp70 Mrna ◽

Shock Response ◽

Attractive Therapeutic Target

Translation elongation factor eEF1A has a well-defined role in protein synthesis. In this study, we demonstrate a new role for eEF1A: it participates in the entire process of the heat shock response (HSR) in mammalian cells from transcription through translation. Upon stress, isoform 1 of eEF1A rapidly activates transcription of HSP70 by recruiting the master regulator HSF1 to its promoter. eEF1A1 then associates with elongating RNA polymerase II and the 3′UTR of HSP70 mRNA, stabilizing it and facilitating its transport from the nucleus to active ribosomes. eEF1A1-depleted cells exhibit severely impaired HSR and compromised thermotolerance. In contrast, tissue-specific isoform 2 of eEF1A does not support HSR. By adjusting transcriptional yield to translational needs, eEF1A1 renders HSR rapid, robust, and highly selective; thus, representing an attractive therapeutic target for numerous conditions associated with disrupted protein homeostasis, ranging from neurodegeneration to cancer.

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MED12 interacts with the heat shock transcription factor HSF1 and recruits CDK8 to promote the heat shock response in mammalian cells

FEBS Letters ◽

10.1002/1873-3468.14139 ◽

2021 ◽

Author(s):

Pratibha Srivastava ◽

Ryosuke Takii ◽

Mariko Okada ◽

Mitsuaki Fujimoto ◽

Akira Nakai

Keyword(s):

Transcription Factor ◽

Heat Shock ◽

Heat Shock Response ◽

Mammalian Cells ◽

Heat Shock Transcription Factor ◽

Shock Response

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Identification of heat-inducible long noncoding RNA MALAT1-containing novel nuclear (HiNoCo) body

Journal of Cell Science ◽

10.1242/jcs.253559 ◽

2021 ◽

Author(s):

Rena Onoguchi-Mizutani ◽

Yoshitaka Kirikae ◽

Yoko Ogura ◽

Tony Gutschner ◽

Sven Diederichs ◽

...

Keyword(s):

Heat Shock ◽

Heat Shock Response ◽

Long Noncoding Rna ◽

Mammalian Cells ◽

Noncoding Rna ◽

Heat Shock Factor ◽

Nuclear Bodies ◽

Cajal Body ◽

Heat Shock Factor 1 ◽

Shock Response

The heat shock response is critical for the survival of all organisms. Metastasis-associated long adenocarcinoma transcript 1 (MALAT1) is a long noncoding RNA localized in nuclear speckles, but its physiological role remains elusive. Here, we show that heat shock induces translocation of MALAT1 to a distinct nuclear body named heat shock-inducible noncoding RNA-containing nuclear (HiNoCo) body in mammalian cells. The MALAT1 knockout A549 cells showed reduced proliferation after heat shock. The HiNoCo body, formed by a nearby nuclear speckle, is distinct from any other known nuclear bodies, including the nuclear stress body, Cajal body, germs, paraspeckles, nucleoli, and promyelocytic leukemia body. The formation of HiNoCo body is reversible and independent of heat shock factor 1, the master transcription regulator of the heat shock response. Our results suggest the HiNoCo body participates in heat shock factor 1-independent heat shock responses in mammalian cells.

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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 ◽

Heat Shock Transcription Factor ◽

Genome Wide ◽

Shock Response ◽

Inducible Genes ◽

Transcription Factor 1

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.

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ATF1 Modulates the Heat Shock Response by Regulating the Stress-Inducible Heat Shock Factor 1 Transcription Complex

Molecular and Cellular Biology ◽

10.1128/mcb.00754-14 ◽

2014 ◽

Vol 35 (1) ◽

pp. 11-25 ◽

Cited By ~ 28

Author(s):

Ryosuke Takii ◽

Mitsuaki Fujimoto ◽

Ke Tan ◽

Eiichi Takaki ◽

Naoki Hayashida ◽

...

Keyword(s):

Heat Shock ◽

Heat Shock Response ◽

Mammalian Cells ◽

Target Genes ◽

Acute Stress ◽

Heat Shock Factor ◽

Metabolic Adaptation ◽

Heat Shock Factor 1 ◽

Creb Binding Protein ◽

Shock Response

The heat shock response is an evolutionally conserved adaptive response to high temperatures that controls proteostasis capacity and is regulated mainly by an ancient heat shock factor (HSF). However, the regulation of target genes by the stress-inducible HSF1 transcription complex has not yet been examined in detail in mammalian cells. In the present study, we demonstrated that HSF1 interacted with members of the ATF1/CREB family involved in metabolic homeostasis and recruited them on theHSP70promoter in response to heat shock. The HSF1 transcription complex, including the chromatin-remodeling factor BRG1 and lysine acetyltransferases p300 and CREB-binding protein (CBP), was formed in a manner that was dependent on the phosphorylation of ATF1. ATF1-BRG1 promoted the establishment of an active chromatin state andHSP70expression during heat shock, whereas ATF1-p300/CBP accelerated the shutdown of HSF1 DNA-binding activity during recovery from acute stress, possibly through the acetylation of HSF1. Furthermore, ATF1 markedly affected the resistance to heat shock. These results revealed the unanticipated complexity of the primitive heat shock response mechanism, which is connected to metabolic adaptation.

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