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

1984 ◽  
Vol 4 (12) ◽  
pp. 2573-2579 ◽  
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.

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

1984 ◽  
Vol 4 (12) ◽  
pp. 2573-2579
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.

A family of related proteins is encoded by the major Drosophila heat shock gene family

1982 ◽  
Vol 2 (3) ◽  
pp. 286-292
Author(s):  
S C Wadsworth
Keyword(s):  
Heat Shock ◽  
Heat Shock Proteins ◽  
Heat Shock Protein ◽  
Sodium Dodecyl ◽  
Heat Shock Gene ◽  
Partial Digestion ◽  
Shock Gene ◽  
Shock Proteins

At least four proteins of 70,000 to 75,000 molecular weight (70-75K) were synthesized from mRNA which hybridized with a cloned heat shock gene previously shown to be localized to the 87A and 87C heat shock puff sites. These in vitro-synthesized proteins were indistinguishable from in vivo-synthesized heat shock-induced proteins when analyzed on sodium dodecyl sulfate-polyacrylamide gels. A comparison of the pattern of this group of proteins synthesized in vivo during a 5-min pulse or during continuous labeling indicates that the 72-75K proteins are probably not kinetic precursors to the major 70K heat shock protein. Partial digestion products generated with V8 protease indicated that the 70-75K heat shock proteins are closely related, but that there are clear differences between them. The partial digestion patterns obtained from heat shock proteins from the Kc cell line and from the Oregon R strain of Drosophila melanogaster are very similar. Genetic analysis of the patterns of 70-75K heat shock protein synthesis indicated that the genes encoding at least two of the three 72-75K heat shock proteins are located outside of the major 87A and 87C puff sites.

scholarly journals Three light-inducible heat shock genes of Chlamydomonas reinhardtii.

1989 ◽  
Vol 9 (9) ◽  
pp. 3911-3918 ◽  
Author(s):  
E D von Gromoff ◽  
U Treier ◽  
C F Beck
Keyword(s):  
Thermal Stress ◽  
Elevated Temperature ◽  
Heat Shock ◽  
Chlamydomonas Reinhardtii ◽  
Heat Shock Gene ◽  
Mrna Levels ◽  
Heat Shock Genes ◽  
Drastic Increase ◽  
Shock Gene

Genomic clones representing three Chlamydomonas reinhardtii genes homologous to the Drosophila hsp70 heat shock gene were isolated. The mRNAs of genes hsp68, hsp70, and hsp80 could be translated in vitro into proteins of Mr 68,000, 70,000, and 80,000, respectively. Transcription of these genes increased dramatically upon heat shock, and the corresponding mRNAs rapidly accumulated, reaching a peak at around 30 min after a shift to the elevated temperature. Light also induced the accumulation of the mRNAs encoded by these heat shock genes. A shift of dark-grown cells to light resulted in a drastic increase in mRNA levels, which reached a maximum at around 1 h after the shift. Thus, in Chlamydomonas, expression of hsp70-homologous heat shock genes appears to be regulated by thermal stress and light.

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

1984 ◽  
Vol 4 (8) ◽  
pp. 1454-1459
Author(s):  
M S Ellwood ◽  
E A Craig
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Heat Shock ◽  
Heat Shock Gene ◽  
Control Analysis ◽  
Coding Region ◽  
Protein Coding ◽  
Shock Gene ◽  
A Minor ◽  
Flanking Regions ◽  
Beta Galactosidase

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.

Three light-inducible heat shock genes of Chlamydomonas reinhardtii

1989 ◽  
Vol 9 (9) ◽  
pp. 3911-3918
Author(s):  
E D von Gromoff ◽  
U Treier ◽  
C F Beck
Keyword(s):  
Thermal Stress ◽  
Elevated Temperature ◽  
Heat Shock ◽  
Chlamydomonas Reinhardtii ◽  
Heat Shock Gene ◽  
Mrna Levels ◽  
Heat Shock Genes ◽  
Drastic Increase ◽  
Shock Gene

Genomic clones representing three Chlamydomonas reinhardtii genes homologous to the Drosophila hsp70 heat shock gene were isolated. The mRNAs of genes hsp68, hsp70, and hsp80 could be translated in vitro into proteins of Mr 68,000, 70,000, and 80,000, respectively. Transcription of these genes increased dramatically upon heat shock, and the corresponding mRNAs rapidly accumulated, reaching a peak at around 30 min after a shift to the elevated temperature. Light also induced the accumulation of the mRNAs encoded by these heat shock genes. A shift of dark-grown cells to light resulted in a drastic increase in mRNA levels, which reached a maximum at around 1 h after the shift. Thus, in Chlamydomonas, expression of hsp70-homologous heat shock genes appears to be regulated by thermal stress and light.

scholarly journals The Heat Shock Protein YbeY Is Required for Optimal Activity of the 30S Ribosomal Subunit

2010 ◽  
Vol 192 (18) ◽  
pp. 4592-4596 ◽  
Author(s):  
Aviram Rasouly ◽  
Chen Davidovich ◽  
Eliora Z. Ron
Keyword(s):  
Heat Shock ◽  
Ribosomal Subunit ◽  
Heat Shock Gene ◽  
Wild Type ◽  
Lower Efficiency ◽  
Ribosomal Subunits ◽  
Translation Machinery ◽  
Shock Gene

ABSTRACT The highly conserved bacterial ybeY gene is a heat shock gene whose function is not fully understood. Previously, we showed that the YbeY protein is involved in protein synthesis, as Escherichia coli mutants with ybeY deleted exhibit severe translational defects in vivo. Here we show that the in vitro activity of the translation machinery of ybeY deletion mutants is significantly lower than that of the wild type. We also show that the lower efficiency of the translation machinery is due to impaired 30S small ribosomal subunits.

scholarly journals Role of DnaK in In Vitro and In Vivo Expression of Virulence Factors of Vibrio cholerae

1999 ◽  
Vol 67 (3) ◽  
pp. 1025-1033 ◽  
Author(s):  
Sabyasachi Chakrabarti ◽  
Nilanjan Sengupta ◽  
Rukhsana Chowdhury
Keyword(s):  
Heat Shock ◽  
Vibrio Cholerae ◽  
Virulence Factors ◽  
Heat Shock Gene ◽  
Insertion Mutant ◽  
In Vivo Expression ◽  
Shock Gene

ABSTRACT The dnaK gene of Vibrio cholerae was cloned, sequenced, and used to construct a dnaK insertion mutant which was then used to examine the role of DnaK in expression of the major virulence factors of this important human pathogen. The central regulator of several virulence genes of V. choleraeis ToxR, a transmembrane DNA binding protein. The V. cholerae dnaK mutant grown in standard laboratory medium exhibited phenotypes characteristic of cells deficient in ToxR activity. Using Northern blot analysis and toxR transcriptional fusions, we demonstrated a reduction in expression of the toxR gene in the dnaK mutant strain together with a concomitant increase in expression of a htpG-like heat shock gene that is located immediately upstream and is divergently transcribed fromtoxR. This may be due to increased heat shock induction in the dnaK mutant. In vivo, however, although expression from heat shock promoters in the dnaK mutant was similar to that observed in vitro, expression of both toxR andhtpG was comparable to that by the parental strain. In both strains, in vivo expression of toxR was significantly higher than that observed in vitro, but no reciprocal decrease inhtpG expression was observed. These results suggest that the modulation of toxR expression in vivo may be different from that observed in vitro.

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

1984 ◽  
Vol 4 (8) ◽  
pp. 1454-1459 ◽  
Author(s):  
M S Ellwood ◽  
E A Craig
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Heat Shock ◽  
Heat Shock Gene ◽  
Control Analysis ◽  
Coding Region ◽  
Protein Coding ◽  
Shock Gene ◽  
A Minor ◽  
Flanking Regions ◽  
Beta Galactosidase

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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