Upstream sequences required for efficient expression of a soybean heat shock gene

1986 ◽  
Vol 6 (2) ◽  
pp. 559-565
Author(s):  
W B Gurley ◽  
E Czarnecka ◽  
R T Nagao ◽  
J L Key
Keyword(s):  
Heat Shock ◽  
Dna Sequences ◽  
Small Heat Shock Protein ◽  
Heat Shock Gene ◽  
Base Pairs ◽  
Basal Transcription ◽  
S1 Nuclease ◽  
Soybean Seedlings ◽  
Flanking Sequences ◽  
Shock Gene

A soybean gene (Gmhsp17.5-E) encoding a small heat shock protein was introduced into primary sunflower tumors via T-DNA-mediated transformation. RNA blot hybridizations and S1-nuclease hybrid protection studies indicated that the heat shock gene containing 3.25 kilobases of 5'-flanking sequences was strongly transcribed in a thermoinducible (40 degrees C) manner. Transcriptional induction also occurred to a lesser extent upon treatment of whole tumors with sodium arsenite and CdCl2. Basal (26 degrees C) transcription was not detected in soybean seedlings, but it was quite evident in transformed tumor tissue. A 5' deletion to -1,175 base pairs with respect to the CAP site had no effect on the levels of thermoinducible transcription, but it resulted in a large increase in basal transcription. Further removal of DNA sequences (including the TATA-distal heat shock consensus element) to -95 base pairs reduced thermoinducible transcription by 95% and also greatly decreased basal transcription. The termini of the Gmhsp17.5-E RNA in the tumor were generally the same as those present in soybean RNA, with the exception of several additional 3' termini.

scholarly journals Upstream sequences required for efficient expression of a soybean heat shock gene.

1986 ◽  
Vol 6 (2) ◽  
pp. 559-565 ◽  
Author(s):  
W B Gurley ◽  
E Czarnecka ◽  
R T Nagao ◽  
J L Key
Keyword(s):  
Heat Shock ◽  
Dna Sequences ◽  
Small Heat Shock Protein ◽  
Heat Shock Gene ◽  
Base Pairs ◽  
Basal Transcription ◽  
S1 Nuclease ◽  
Soybean Seedlings ◽  
Flanking Sequences ◽  
Shock Gene

A soybean gene (Gmhsp17.5-E) encoding a small heat shock protein was introduced into primary sunflower tumors via T-DNA-mediated transformation. RNA blot hybridizations and S1-nuclease hybrid protection studies indicated that the heat shock gene containing 3.25 kilobases of 5'-flanking sequences was strongly transcribed in a thermoinducible (40 degrees C) manner. Transcriptional induction also occurred to a lesser extent upon treatment of whole tumors with sodium arsenite and CdCl2. Basal (26 degrees C) transcription was not detected in soybean seedlings, but it was quite evident in transformed tumor tissue. A 5' deletion to -1,175 base pairs with respect to the CAP site had no effect on the levels of thermoinducible transcription, but it resulted in a large increase in basal transcription. Further removal of DNA sequences (including the TATA-distal heat shock consensus element) to -95 base pairs reduced thermoinducible transcription by 95% and also greatly decreased basal transcription. The termini of the Gmhsp17.5-E RNA in the tumor were generally the same as those present in soybean RNA, with the exception of several additional 3' termini.

scholarly journals Sequences involved in temperature and ecdysterone-induced transcription are located in separate regions of a Drosophila melanogaster heat shock gene.

1986 ◽  
Vol 6 (2) ◽  
pp. 663-673 ◽  
Author(s):  
E Hoffman ◽  
V Corces
Keyword(s):  
Drosophila Melanogaster ◽  
Heat Shock ◽  
Dna Sequences ◽  
Germ Line ◽  
Initiation Site ◽  
P Element ◽  
Heat Shock Gene ◽  
Base Pairs ◽  
Consensus Sequences ◽  
Shock Gene

The transcriptional regulation of the Drosophila melanogaster hsp27 (also called hsp28) gene was studied by introducing altered genes into the germ line by P element-mediated transformation. DNA sequences upstream of the gene were defined with respect to their effect on steroid hormone-induced and heat-induced transcription. These two types of control were found to be separable; the sequences responsible for 80% of heat-induced expression were located more than 1.1 kilobases upstream of the RNA initiation site, while the sequences responsible for the majority of ecdysterone induction were positioned downstream of the site at -227 base pairs. We have determined the DNA sequence of the intergenic region separating hsp23 and hsp27 and have located putative heat shock and ecdysterone consensus sequences. Our results indicate that the heat shock promoter of the hsp27 gene is organized quite differently from that of hsp70.

Sequences involved in temperature and ecdysterone-induced transcription are located in separate regions of a Drosophila melanogaster heat shock gene

1986 ◽  
Vol 6 (2) ◽  
pp. 663-673
Author(s):  
E Hoffman ◽  
V Corces
Keyword(s):  
Drosophila Melanogaster ◽  
Heat Shock ◽  
Dna Sequences ◽  
Germ Line ◽  
Initiation Site ◽  
P Element ◽  
Heat Shock Gene ◽  
Base Pairs ◽  
Consensus Sequences ◽  
Shock Gene

The transcriptional regulation of the Drosophila melanogaster hsp27 (also called hsp28) gene was studied by introducing altered genes into the germ line by P element-mediated transformation. DNA sequences upstream of the gene were defined with respect to their effect on steroid hormone-induced and heat-induced transcription. These two types of control were found to be separable; the sequences responsible for 80% of heat-induced expression were located more than 1.1 kilobases upstream of the RNA initiation site, while the sequences responsible for the majority of ecdysterone induction were positioned downstream of the site at -227 base pairs. We have determined the DNA sequence of the intergenic region separating hsp23 and hsp27 and have located putative heat shock and ecdysterone consensus sequences. Our results indicate that the heat shock promoter of the hsp27 gene is organized quite differently from that of hsp70.

Regulatory domains of the Gmhsp17.5-E heat shock promoter of soybean

1989 ◽  
Vol 9 (8) ◽  
pp. 3457-3463
Author(s):  
E Czarnecka ◽  
J L Key ◽  
W B Gurley
Keyword(s):  
Heat Shock ◽  
Promoter Activity ◽  
Heat Shock Gene ◽  
Base Pairs ◽  
Transgenic Expression ◽  
Cis Acting ◽  
Regulatory Domains ◽  
Shock Gene ◽  
Tata Motif

Promoter domains required for in vivo transcriptional expression of soybean heat shock gene Gmhsp17.5-E were identified by insertion-deletion mutagenesis with transgenic expression monitored in Agrobacterium tumefaciens-incited tumors of sunflower. Removal of the TATA-distal domain from position -1175 to position -259 had little effect on overall activity. The four regions contributing to promoter activity identified by this study all map within 244 base pairs from the start of transcription. The most distal cis-acting element of major significance was located from -244 to -179 and contains a conserved TATA-dyad motif centered at -220. Sequences from -179 to -40 comprise the TATA-proximal domain and include an AT-rich region and two sites containing heat shock consensus elements (HSEs). Deletion of the HSE centered at -93 (site 2) severely reduced transcriptional activity. Heat-inducible expression was also eliminated by internal deletion of either the TATA motif or the overlapping HSEs at site 1, indicating that each of these regions is also a major determinant of promoter activity.

scholarly journals A New Heat Shock Gene, agsA, Which Encodes a Small Chaperone Involved in Suppressing Protein Aggregation in Salmonella enterica Serovar Typhimurium

2003 ◽  
Vol 185 (21) ◽  
pp. 6331-6339 ◽  
Author(s):  
Toshifumi Tomoyasu ◽  
Akiko Takaya ◽  
Tomomi Sasaki ◽  
Takahiro Nagase ◽  
Reiko Kikuno ◽  
...  
Keyword(s):  
Heat Shock ◽  
High Temperatures ◽  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Transcriptional Start Site ◽  
Small Heat Shock Protein ◽  
Heat Shock Gene ◽  
Evolutionary Stage ◽  
Serovar Typhimurium ◽  
Shock Gene

ABSTRACT We discovered a novel small heat shock protein (sHsp) named AgsA (aggregation-suppressing protein) in the thermally aggregated fraction from a Salmonella enterica serovar Typhimurium dnaK-null strain. The −10 and −35 regions upstream of the transcriptional start site of the agsA gene are characteristic of σ32- and σ72-dependent promoters. AgsA was strongly induced by high temperatures. The similarity between AgsA and the other two sHsps of Salmonella serovar Typhimurium, IbpA and IbpB, is rather low (around 30% amino acid sequence identity). Phylogenetic analysis suggested that AgsA arose from an ancient gene duplication or amplification at an early evolutionary stage of gram-negative bacteria. Here we show that overproduction of AgsA partially complements the ΔdnaK52 thermosensitive phenotype and reduces the amount of heat-aggregated proteins in both ΔdnaK52 and ΔrpoH mutants of Escherichia coli. These data suggest that AgsA is an effective chaperone capable of preventing aggregation of nonnative proteins and maintaining them in a state competent for refolding in Salmonella serovar Typhimurium at high temperatures.

scholarly journals Regulated transcription of the genes for actin and heat-shock proteins in culture drosophila cells

1981 ◽  
Vol 88 (2) ◽  
pp. 323-328 ◽  
Author(s):  
RC Findly ◽  
T Pederson
Keyword(s):  
Heat Shock ◽  
Heat Shock Proteins ◽  
Gene Transcription ◽  
Dna Sequences ◽  
Heat Shock Gene ◽  
Nuclear Rna ◽  
Gene Transcripts ◽  
Shock Gene ◽  
Shock Proteins ◽  
Drosophila Cells

The transcription of three specific genes has been examined in heat-shocked drosophila cells by hybridizing pulse-labeled nuclear RNA with cloned DNA sequences. Actin gene transcription is rapidly and profoundly suppressed upon heat shock but returns to near- normal levels after cells are placed back at their normal culture temperature (23 degrees C). Conversely, the transcription of genes coding from 70,000- and 26,000-dalton heat- shock proteins increases dramatically and with extraordinary rapidity (60 s) after heat shock. The temporal patterns of 70,000- and 26,000-dalton heat-shock gene transcription are nearly superimposable, indicating that, although they are closely linked cytologically, these genes are nevertheless tightly coregulated. The abundance of heat- shock gene transcripts reaches remarkable levels, e.g., 70,000-dalton heat-shock gene transcripts account for 2-3 percent of the nuclear RNA labeled during the first 30 min of heat shock. When heat-shocked cells are returned to 25 degrees C, the rates of transcription of the heat-shock genes fall back to the low levels characteristic of untreated cells. To confirm the low level of heat-shock gene transcription in normal cells, nuclear RNA was purified from unlabeled (and otherwise unhandled) 25 degrees C cells, end-labeled in vitro with (32)P, and hybridized to cloned heat-shock DNA sequences. These and other data establish that the genes for 70,000- and 26,000-dalton heat-shock proteins in culture drosophila cells are active at 25 degrees C, and that their rate of transcription is greatly accelerated upon heat shock rather than being activated from a true "off" state. The rapidity, magnitude, and reversibility of the shifts in actin and heat-shock gene transcription constitute compelling advantages for the use of cultured drosophila cells in studying the transcriptional regulation of eukaryotic genes, including one related to the cytoskeleton.

Characterization of expressed meiotic prophase repeat transcript clones of Lilium: meiosis-specific expression, relatedness, and affinities to small heat shock protein genes

Genome ◽  
1990 ◽  
Vol 33 (1) ◽  
pp. 68-79 ◽  
Author(s):  
Robert A. Bouchard
Keyword(s):  
Heat Shock ◽  
Meiotic Prophase ◽  
Higher Plants ◽  
Small Heat Shock Protein ◽  
Heat Shock Gene ◽  
Cdna Clones ◽  
Restriction Maps ◽  
Specific Expression ◽  
Shock Gene

The inserts of plasmid cDNA clones for transcripts showing meiotic prophase specific expression show cross reassociation to varying degrees of intensity with one another. These clones were recovered from a cDNA library made from Lilium microsporocyte poly(A)+ RNA. RNA-dot and Northern-blot analyses indicate that these clones represent transcripts specific to the meiotic prophase interval in microsporocytes. The transcripts appear to constitute the most abundant class of meiosis-specific poly(A)+ RNAs. At least two subgroups can be distinguished by examining cloned transcripts from genes of this expressed meiotic prophase repeat (EMPR) sequence family. Members of each subgroup have similar although not identical restriction maps and show relatively high but varying fidelities of DNA cross reassociation between members. However, consensus restriction maps of the two subgroups are largely dissimilar and, except at low stringencies, cross reassociation is readily detected only at restriction fragments from a particular conserved internal segment. The DNA sequence of a representative EMPR clone has been determined, and the inferred peptide product has been found to show extensive sequence homology to that of a small heat-shock gene of Glycine max, particularly in the conserved region. Alignment of the sequences for the conserved regions of two EMPR subgroup representatives with the soybean sequence suggests that selection has acted to conserve similar blocks of amino acids in this area. These observations suggest that a major portion of the transcripts produced during the apparently unrelated processes of meiosis and heat shock in higher plants are derived from related gene sequences encoding similar products.Key words: meiosis, transcription, specific cDNA, heat-shock mRNA.

scholarly journals Regulatory domains of the Gmhsp17.5-E heat shock promoter of soybean.

1989 ◽  
Vol 9 (8) ◽  
pp. 3457-3463 ◽  
Author(s):  
E Czarnecka ◽  
J L Key ◽  
W B Gurley
Keyword(s):  
Heat Shock ◽  
Promoter Activity ◽  
Heat Shock Gene ◽  
Base Pairs ◽  
Transgenic Expression ◽  
Cis Acting ◽  
Regulatory Domains ◽  
Shock Gene ◽  
Tata Motif

Promoter domains required for in vivo transcriptional expression of soybean heat shock gene Gmhsp17.5-E were identified by insertion-deletion mutagenesis with transgenic expression monitored in Agrobacterium tumefaciens-incited tumors of sunflower. Removal of the TATA-distal domain from position -1175 to position -259 had little effect on overall activity. The four regions contributing to promoter activity identified by this study all map within 244 base pairs from the start of transcription. The most distal cis-acting element of major significance was located from -244 to -179 and contains a conserved TATA-dyad motif centered at -220. Sequences from -179 to -40 comprise the TATA-proximal domain and include an AT-rich region and two sites containing heat shock consensus elements (HSEs). Deletion of the HSE centered at -93 (site 2) severely reduced transcriptional activity. Heat-inducible expression was also eliminated by internal deletion of either the TATA motif or the overlapping HSEs at site 1, indicating that each of these regions is also a major determinant of promoter activity.

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