Regulation of heat-shock genes: a DNA sequence upstream of Drosophila hsp70 genes is essential for their induction in monkey cells.

Mirault, Marc-Édouard; Southgate, Richard; Delwart, Eric

doi:10.1002/j.1460-2075.1982.tb00025.x

Cited by 177 publications

(89 citation statements)

References 42 publications

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“…The HSE functions efficiently upstream of heterologous genes when reintroduced into Drosophila, Xenopus, murine, and human cells (12,(39)(40)(41). Sequences with homology to HSEs are also located upstream of Xenopus and soybean heat shock genes (42,43).…”

Section: Discussionmentioning

confidence: 99%

“…One class of inducible genes with complex regulation includes the heat shock or stress-induced genes. The expression of heat shock genes is induced by a wide range of stimuli, including heat shock, heavy metals, cessation ofanoxia, inhibitors of energy metabolism, amino acid analogues, infection by adenovirus 5 or simian virus 40 (SV40), stimulation with serum, and development (22)(23)(24)(25).…”

mentioning

confidence: 99%

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Human HSP70 promoter contains at least two distinct regulatory domains.

Wu¹,

Kingston²,

Morimoto³

1986

Proc. Natl. Acad. Sci. U.S.A.

298

200

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The expression of the human HSP70 gene is induced by a wide range of physiological stresses, including exposure to heat shock and heavy metals, or under nonstress conditions, such as in response to serum stimulation. We have previously demonstrated that in either case the regulated expression is at the primary level of transcription. To determine whether transcription is mediated through a single or multiple genetic elements, we have dissected the sequences upstream of the transcription start site of the human HSP70 gene by constructing chimeric genes retaining variable amounts of 5' flanking regions fused to the bacterial gene encoding chloramphenicol acetyltransferase. Transcription from the chimeric genes was determined by S1 nuclease analysis of separate stable transfectants. The sequences required for heat shock and cadmium induction lie between -107 and -68. Within this region is the sequence CTGGAATAT-TCCCG, which is identical in 12/14 positions with the heat shock element of Drosophila heat shock genes, and a separate sequence, CGNCCCGG, which is homologous to the core of the human metallothionein II metal-responsive element. The sequences required for serum-stimulated transcription are distinct from the heat shock element. The sequence CCAAT at -68 is required for high levels ofcorrectly initiated transcripts, and a purine-rich sequence, GAAGGGAAAAG, at -58 is required for serum stimulation. The human HSP70 promoter contains at least two regulatory domains-a distal domain responsive to heat shock or cadmium and a proximal domain responsive to stimulation by serum.Sequences defining eukaryotic promoters can be separated into three functional classes. The first class is enhancer elements: sequences that function independent of location or orientation and, in some cases, confer tissue specificity (1). The second class is elements that confer the basal activity of promoters and determine the start site of initiation; this class includes the "TATA box" (2-4) and the "CAAT homology" (5-9). These elements generally function in close proximity to the transcriptional start site. It is not clear whether these elements can play a role in regulation oftranscription. A third class of elements, regulatory elements, confers inducible regulation in response to a specific stimulus.Inducible promoters, such as those regulating metallothionein, mouse mammary tumor virus, interferon, and heat shock genes, have been particularly useful in studying rapid changes in transcription. Studies localizing regulatory elements responsive to specific inducers have identified the heat shock elements (HSE) for heat shock genes (10-13), metal-responsive elements (MRE) for metallothionein genes (14, 15), glucocorticoid-responsive elements (GRE) for mouse mammary tumor virus genes (16-18), and interferonregulatory elements (IRE) for the viral or poly(dI-dC) induction of a-and (3-interferon genes (19-21). One class of inducible genes with complex regulation includes the heat shock or stress-induced genes. The expression of heat shoc...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Human HSP70 promoter contains at least two distinct regulatory domains.

Wu¹,

Kingston²,

Morimoto³

1986

Proc. Natl. Acad. Sci. U.S.A.

298

200

View full text Add to dashboard Cite

show abstract

“…Genetic evidence from 87A deficiencies limits the maximum amount of 5' flanking sequence required for regulated expression in vivo to 479 nucleotides (43). In vitro generated deletions assayed in heterologous systems suggest that only about 70 nucleotides of flanking sequence are required for heat induction (44)(45)(46)(47). The (46).…”

Section: Discussionmentioning

confidence: 99%

Localization of the hsp83 transcript within a 3292 nucleotide sequence from the 63B heat shock locus ofD. melanogaster

Hackett¹,

Lis²

1983

Nucl Acids Res

109

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We have determined the complete nucleotide sequence of a 3292 bp cloned segment derived from the 63B heat shock cytogenetic locus of D. melanogaster. Within this segment we have positioned the start of transcription and RNA splice sites of the unique gene that encodes the 83,000 d heat shock polypeptide (hsp83 gene) by SI mapping and synthesis of cDNA from restriction fragment primed mRNA. The sequence begins at a point 879 bp upstream from the transcription start and includes the 149 bp nontranslated first exon, the 1139 bp intron and extends 1125 bp into the protein coding region. These data identify a single open translation reading frame for the first 375 amino acids of the 83,000 d polypeptide, beginning with the first ATG codon located at the 3' intron-exon junction. We discuss and demonstrate the use of E. coli exonuclease III generated single-strand DNA probes as an alternative to strand separation for S1 mapping of mRNA. We also use homology search criteria based upon known protein-DNA binding sites to compare our hsp83 sequence with other sequenced Drosophila heat shock genes. These comparisons indicate that a large region of approximately 80 bp centered around the transcription initiation point of the hsp83 gene shares only a 31% homology with the corresponding region of the hsp70 gene, whereas the hsp22, 23, 26, and 27 genes share a 54% homology with hsp70 in this region. The lower homology of the hsp83 gene is consistent with the deviant nature of this heat shock gene.

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“…Based on all of these results, it seems most likely that different factors are involved in ElA control; this was borne out by our experiments with the same hsp7O gene. A heat shock control element (30,39) is found at position -110 relative to the transcription initiation site of the human hsp7O gene, and plasmids that contain this sequence are both ElA and heat shock inducible. Deletion of these sequences, leaving only 77 nucleotides of upstream sequence, eliminates the heat shock response but leaves the ElA inducibility intact (M. C.…”

Section: Cellsmentioning

confidence: 99%

Selective induction of human heat shock gene transcription by the adenovirus E1A gene products, including the 12S E1A product.

Simon¹,

Kitchener²,

Kao³

et al. 1987

Mol. Cell. Biol.

111

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We have previously shown that the human 70-kilodalton heat shock protein gene (hsp7O) is induced by the adenovirus ElA gene product and during the S-G2 phase of the cell cycle. In this study, we investigated the effect of ElA on the expression of other human hsp genes. A gene encoding one form of the hsp89 protein (hsp89a) was activated during an adenovirus infection with kinetics similar to those of activation of hsp7O. The induction required a functional ElA gene. However, the hsp89 transcript was not cell cycle regulated. Genes encoding another form of hsp89 and the hsp27 protein were not induced by ElA or during the cell cycle. Further examination of hsp7O expression revealed a greater complexity than previously seen. Si nuclease analysis using an hsp70 cDNA as well as a distinct hsp7O genomic clone demonstrated three related hsp70 transcripts; two were induced by ElA, and one was not. Both of the ElA-inducible genes were regulated during the cell cycle. All three were induced by heat shock. These results suggest common aspects of control among certain members of this family of cellular genes distinct from heat shock control. Finally, using viruses that express the individual ElA proteins, we found that the hsp70 gene is induced by the 12S and the 13S ElA products. The efficiency of induction by the 12S product was somewhat less than that by the 13S product but only by a factor of less than 2. This is in contrast to the induction of early viral genes, for which the 13S product is considerably more efficient than the 12S product.

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Regulation of heat-shock genes: a DNA sequence upstream of Drosophila hsp70 genes is essential for their induction in monkey cells.

Cited by 177 publications

References 42 publications

Human HSP70 promoter contains at least two distinct regulatory domains.

Human HSP70 promoter contains at least two distinct regulatory domains.

Localization of the hsp83 transcript within a 3292 nucleotide sequence from the 63B heat shock locus ofD. melanogaster

Selective induction of human heat shock gene transcription by the adenovirus E1A gene products, including the 12S E1A product.

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