Differential regulation of the 70K heat shock gene and related genes in Saccharomyces cerevisiae

Saccharomyces cerevisiae contains a family of genes related to Hsp70, the major heat shock gene of Drosophila melanogaster. The transcription of three of these genes, which show no conservation of sequences 5' to the protein-coding region, was analyzed. The 5' flanking regions from the three genes were fused to the Escherichia coli beta-galactosidase structural gene and introduced into yeasts on multicopy plasmids, putting the beta-galactosidase production under yeast promoter control. Analysis of beta-galactosidase mRNA and protein production in these transformed strains revealed that transcription from the three promoters is differentially regulated. The number of transcripts from one promoter is vastly increased for a brief period after heat shock, whereas mRNA from another declines. Transcripts from a third gene are slightly enhanced upon heat shock; however, multiple 5' ends of the mRNA are found, and a minor species increases in amount after heat shock. Transcription of these promoters in their native state on the chromosome appears to be modulated in the same manner.

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Stimuli that induce a yeast heat shock gene fused to beta-galactosidase.

Molecular and Cellular Biology ◽

10.1128/mcb.4.12.2573 ◽

1984 ◽

Vol 4 (12) ◽

pp. 2573-2579 ◽

Cited By ~ 13

Author(s):

C Brazzell ◽

T D Ingolia

Keyword(s):

Heat Shock ◽

Heat Shock Gene ◽

Enzyme Assays ◽

Lacz Gene ◽

Directed Synthesis ◽

Shock Gene ◽

Chemical Stimuli ◽

Physical And Chemical ◽

Beta Galactosidase

Saccharomyces cerevisiae contain a multigene family related to the Drosophila heat shock gene hsp70. Two members of this family, YG100 and YG101, have been previously characterized (Ingolia et al., Mol. Cell. Biol. 2:1388-1398, 1982), and only YG100 was found to have elevated levels of transcription after heat shock. The yeast hsp70 genes contained on YG100 and YG101 were truncated and fused to the Escherichia coli lacZ gene contained on pMC1587 (Casadaban et al., Methods Enzymol. 100:283-308, 1983). The resulting plasmids directed synthesis of the beta-galactosidase gene as measured by in vitro enzyme assays and by colorimetric assays on plates. The expression level from the YG101 gene was constant under all the conditions tested, whereas expression driven by the YG100 gene could be induced over 50-fold. Other stimuli besides heat, including recovery from anoxia and high cell density, were found to strongly induce YG100 gene expression. Most physical and chemical stimuli tested, including UV irradiation, zymolyase treatment, and ethanol, did not stimulate expression of this heat shock gene.

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Transcriptional regulation of an hsp70 heat shock gene in the yeast Saccharomyces cerevisiae

Molecular and Cellular Biology ◽

10.1128/mcb.7.5.1906-1916.1987 ◽

1987 ◽

Vol 7 (5) ◽

pp. 1906-1916

Author(s):

M R Slater ◽

E A Craig

Keyword(s):

Saccharomyces Cerevisiae ◽

Heat Shock ◽

Promoter Region ◽

Inducible Expression ◽

Proximal Promoter ◽

Heat Shock Gene ◽

Heat Shock Element ◽

Yeast Saccharomyces Cerevisiae ◽

Hybrid Promoter ◽

Shock Gene

The yeast Saccharomyces cerevisiae contains three heat-inducible hsp70 genes. We have characterized the promoter region of the hsp70 heat shock gene YG100, that also displays a basal level of expression. Deletion of the distal region of the promoter resulted in an 80% drop in the basal level of expression without affecting expression after heat shock. Progressive-deletion analysis suggested that sequences necessary for heat-inducible expression are more proximal, within 233 base pairs of the initiation region. The promoter region of YG100 contains multiple elements related to the Drosophila melanogaster heat shock element (HSE; CnnGAAnnT TCnnG). Deletion of a proximal promoter region containing one element, HSE2, eliminated most of the heat-inducible expression of YG100. The upstream activation site (UAS) of the yeast cytochrome c gene (CYC1) can be substituted by a single copy of HSE2 plus its adjoining nucleotides (UASHS). This hybrid promoter displayed a substantial level of expression before heat shock, and the level of expression was elevated eightfold by heat shock. YG100 sequences that flank UASHS inhibited basal expression of UASHS in the hybrid promoter but not its heat-inducible expression. This inhibition of basal UASHS activity suggests that negative regulation is involved in modulating expression of this yeast heat shock gene.

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Transcriptional regulation of an hsp70 heat shock gene in the yeast Saccharomyces cerevisiae.

Molecular and Cellular Biology ◽

10.1128/mcb.7.5.1906 ◽

1987 ◽

Vol 7 (5) ◽

pp. 1906-1916 ◽

Cited By ~ 67

Author(s):

M R Slater ◽

E A Craig

Keyword(s):

Saccharomyces Cerevisiae ◽

Heat Shock ◽

Promoter Region ◽

Inducible Expression ◽

Proximal Promoter ◽

Heat Shock Gene ◽

Heat Shock Element ◽

Yeast Saccharomyces Cerevisiae ◽

Hybrid Promoter ◽

Shock Gene

The yeast Saccharomyces cerevisiae contains three heat-inducible hsp70 genes. We have characterized the promoter region of the hsp70 heat shock gene YG100, that also displays a basal level of expression. Deletion of the distal region of the promoter resulted in an 80% drop in the basal level of expression without affecting expression after heat shock. Progressive-deletion analysis suggested that sequences necessary for heat-inducible expression are more proximal, within 233 base pairs of the initiation region. The promoter region of YG100 contains multiple elements related to the Drosophila melanogaster heat shock element (HSE; CnnGAAnnT TCnnG). Deletion of a proximal promoter region containing one element, HSE2, eliminated most of the heat-inducible expression of YG100. The upstream activation site (UAS) of the yeast cytochrome c gene (CYC1) can be substituted by a single copy of HSE2 plus its adjoining nucleotides (UASHS). This hybrid promoter displayed a substantial level of expression before heat shock, and the level of expression was elevated eightfold by heat shock. YG100 sequences that flank UASHS inhibited basal expression of UASHS in the hybrid promoter but not its heat-inducible expression. This inhibition of basal UASHS activity suggests that negative regulation is involved in modulating expression of this yeast heat shock gene.

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UBI4, the polyubiquitin gene of Saccharomyces cerevisiae, is a heat shock gene that is also subject to catabolite derepression control

MGG Molecular & General Genetics ◽

10.1007/s004380050341 ◽

1997 ◽

Vol 253 (4) ◽

pp. 439-447 ◽

Cited By ~ 22

Author(s):

R. Watt ◽

P. W. Piper

Keyword(s):

Saccharomyces Cerevisiae ◽

Heat Shock ◽

Heat Shock Gene ◽

Polyubiquitin Gene ◽

Shock Gene

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Interdependence of several heat shock gene activations, cyclic AMP decline and changes at the plasma membrane of Saccharomyces cerevisiae

Antonie van Leeuwenhoek ◽

10.1007/bf00548933 ◽

1990 ◽

Vol 58 (3) ◽

pp. 195-201 ◽

Cited By ~ 6

Author(s):

Peter Piper

Keyword(s):

Saccharomyces Cerevisiae ◽

Plasma Membrane ◽

Heat Shock ◽

Cyclic Amp ◽

Heat Shock Gene ◽

Shock Gene

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Requirements for chromatin reassembly during transcriptional downregulation of a heat shock gene in Saccharomyces cerevisiae

FEBS Journal ◽

10.1111/j.1742-4658.2008.06451.x ◽

2008 ◽

Vol 275 (11) ◽

pp. 2956-2964 ◽

Cited By ~ 15

Author(s):

Mette M. Jensen ◽

Marianne S. Christensen ◽

Bjarne Bonven ◽

Torben H. Jensen

Keyword(s):

Saccharomyces Cerevisiae ◽

Heat Shock ◽

Heat Shock Gene ◽

Shock Gene

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A heat shock gene from Saccharomyces cerevisiae encoding a secretory glycoprotein.

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.89.9.3671 ◽

1992 ◽

Vol 89 (9) ◽

pp. 3671-3675 ◽

Cited By ~ 90

Author(s):

P. Russo ◽

N. Kalkkinen ◽

H. Sareneva ◽

J. Paakkola ◽

M. Makarow

Keyword(s):

Saccharomyces Cerevisiae ◽

Heat Shock ◽

Heat Shock Gene ◽

Shock Gene

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The Archaeal Molecular Chaperone Machine: Peculiarities and Paradoxes

Genetics ◽

10.1093/genetics/152.4.1277 ◽

1999 ◽

Vol 152 (4) ◽

pp. 1277-1283

Author(s):

Alberto J L Macario ◽

Everly Conway de Macario

Keyword(s):

Heat Shock ◽

Molecular Chaperone ◽

Molecular Chaperones ◽

Eukaryotic Cell ◽

Heat Shock Gene ◽

Physiological Importance ◽

Shock Gene ◽

Cell Cytosol ◽

High Degree ◽

Flanking Regions

Abstract A major finding within the field of archaea and molecular chaperones has been the demonstration that, while some species have the stress (heat-shock) gene hsp70(dnaK), others do not. This gene encodes Hsp70(DnaK), an essential molecular chaperone in bacteria and eukaryotes. Due to the physiological importance and the high degree of conservation of this protein, its absence in archaeal organisms has raised intriguing questions pertaining to the evolution of the chaperone machine as a whole and that of its components in particular, namely, Hsp70(DnaK), Hsp40(DnaJ), and GrpE. Another archaeal paradox is that the proteins coded by these genes are very similar to bacterial homologs, as if the genes had been received via lateral transfer from bacteria, whereas the upstream flanking regions have no bacterial markers, but instead have typical archaeal promoters, which are like those of eukaryotes. Furthermore, the chaperonin system in all archaea studied to the present, including those that possess a bacterial-like chaperone machine, is similar to that of the eukaryotic-cell cytosol. Thus, two chaperoning systems that are designed to interact with a compatible partner, e.g., the bacterial chaperone machine physiologically interacts with the bacterial but not with the eucaryal chaperonins, coexist in archaeal cells in spite of their apparent functional incompatibility. It is difficult to understand how these hybrid characteristics of the archaeal chaperoning system became established and work, if one bears in mind the classical ideas learned from studying bacteria and eukaryotes. No doubt, archaea are intriguing organisms that offer an opportunity to find novel molecules and mechanisms that will, most likely, enhance our understanding of the stress response and the protein folding and refolding processes in the three phylogenetic domains.

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