Organization of the Drosophila melanogaster hsp70 heat shock regulation unit

1987 ◽  
Vol 7 (3) ◽  
pp. 1055-1062
Author(s):  
J Amin ◽  
R Mestril ◽  
P Schiller ◽  
M Dreano ◽  
R Voellmy
Keyword(s):  
Drosophila Melanogaster ◽  
Heat Shock ◽  
Synergistic Effect ◽  
Consensus Sequence ◽  
Relative Orientation ◽  
Hsp70 Promoter ◽  
Promoter Sequences ◽  
Highly Active ◽  
Sequence Elements ◽  
Five Elements

Expression from the Drosophila melanogaster hsp70 promoter was controlled by a regulatory unit that was composed of two sequence elements that resembled the heat shock consensus sequence. The unit functioned in both orientations and at different distances from downstream promoter sequences. Each element of the unit alone was essentially inactive. Association of two elements resulted in a dramatic increase of transcription from the hsp70 promoter. This synergistic effect was independent of the relative orientation of the elements and, to a large extent, of the distance between them. Duplication of a region containing only one element also yielded a highly active, heat-regulated promoter. Genes with three to five elements were three to four times more active than those with a single regulatory unit.

scholarly journals Organization of the Drosophila melanogaster hsp70 heat shock regulation unit.

1987 ◽  
Vol 7 (3) ◽  
pp. 1055-1062 ◽  
Author(s):  
J Amin ◽  
R Mestril ◽  
P Schiller ◽  
M Dreano ◽  
R Voellmy
Keyword(s):  
Drosophila Melanogaster ◽  
Heat Shock ◽  
Synergistic Effect ◽  
Consensus Sequence ◽  
Relative Orientation ◽  
Hsp70 Promoter ◽  
Promoter Sequences ◽  
Highly Active ◽  
Sequence Elements ◽  
Five Elements

Expression from the Drosophila melanogaster hsp70 promoter was controlled by a regulatory unit that was composed of two sequence elements that resembled the heat shock consensus sequence. The unit functioned in both orientations and at different distances from downstream promoter sequences. Each element of the unit alone was essentially inactive. Association of two elements resulted in a dramatic increase of transcription from the hsp70 promoter. This synergistic effect was independent of the relative orientation of the elements and, to a large extent, of the distance between them. Duplication of a region containing only one element also yielded a highly active, heat-regulated promoter. Genes with three to five elements were three to four times more active than those with a single regulatory unit.

TATA-dependent and TATA-independent function of the basal and heat shock elements of a human hsp70 promoter

1990 ◽  
Vol 10 (4) ◽  
pp. 1319-1328
Author(s):  
J M Greene ◽  
R E Kingston
Keyword(s):  
Heat Shock ◽  
Consensus Sequence ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Hsp70 Promoter ◽  
Heat Shock Element ◽  
Tata Element ◽  
Sequence Elements ◽  
Human Heat Shock Protein ◽  
Human Hsp70

We have characterized the interactions between the TATA element and other sequence elements of a human heat shock protein 70 (hsp70) promoter by a mutational approach. Expression of a distal element of this promoter requires an intact TATA element in human cell lines. The hsp70 TATA element can be functionally replaced for this interaction by TATA elements from the simian virus 40 early and adenovirus EIIa promoters. The TATA element in this promoter therefore both determines the appropriate start site and determines strength by allowing function of the distal element. In contrast, three proximal upstream elements necessary for basal and heat-regulated transcription have no requirement either for a TATA element or for any other proximal element. The behavior of promoters multiply mutant in these proximal elements implies that these elements function independently. We examined the interaction between the heat shock element (HSE) and the TATA element as the distance between the two factor-binding sites was increased. It was necessary to create a mutant HSE with an extended consensus sequence in order for the HSE to function at a distance. Moving this extended HSE 500 bases upstream did not increase its dependence on the TATA element, suggesting that the TATA independence of this element is intrinsic to its function and is not determined by distance from the promoter.

scholarly journals TATA-dependent and TATA-independent function of the basal and heat shock elements of a human hsp70 promoter.

1990 ◽  
Vol 10 (4) ◽  
pp. 1319-1328 ◽  
Author(s):  
J M Greene ◽  
R E Kingston
Keyword(s):  
Heat Shock ◽  
Consensus Sequence ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Hsp70 Promoter ◽  
Heat Shock Element ◽  
Tata Element ◽  
Sequence Elements ◽  
Human Heat Shock Protein ◽  
Human Hsp70

We have characterized the interactions between the TATA element and other sequence elements of a human heat shock protein 70 (hsp70) promoter by a mutational approach. Expression of a distal element of this promoter requires an intact TATA element in human cell lines. The hsp70 TATA element can be functionally replaced for this interaction by TATA elements from the simian virus 40 early and adenovirus EIIa promoters. The TATA element in this promoter therefore both determines the appropriate start site and determines strength by allowing function of the distal element. In contrast, three proximal upstream elements necessary for basal and heat-regulated transcription have no requirement either for a TATA element or for any other proximal element. The behavior of promoters multiply mutant in these proximal elements implies that these elements function independently. We examined the interaction between the heat shock element (HSE) and the TATA element as the distance between the two factor-binding sites was increased. It was necessary to create a mutant HSE with an extended consensus sequence in order for the HSE to function at a distance. Moving this extended HSE 500 bases upstream did not increase its dependence on the TATA element, suggesting that the TATA independence of this element is intrinsic to its function and is not determined by distance from the promoter.

Modifying expression of the engrailed gene of Drosophila melanogaster

Development ◽  
1988 ◽  
Vol 104 (Supplement) ◽  
pp. 85-93 ◽  
Author(s):  
Stephen J. Poole ◽  
Thomas B. Kornberg
Keyword(s):  
Gene Expression ◽  
Drosophila Melanogaster ◽  
Heat Shock ◽  
P Element ◽  
Drosophila Embryo ◽  
Hsp70 Promoter ◽  
Normal Period ◽  
In Cells

The engrailed gene is required for segmentation of the Drosophila embryo and is expressed in cells constituting the posterior developmental compartments. In mutant embryos lacking engrailed function, portions of the cuticular pattern in each segment are deleted, resulting in fusion of adjacent denticle bands. Using P-element-mediated transposition, we generated flies that express the engrailed gene under the control of an hsp70 promoter, and found that ectopic, heat-shock-induced, engrailed expression caused pattern defects similar to those in embryos lacking engrailed function. Sensitivity to heat shock was only during the cellular blastoderm and early gastrulation periods. This window of sensitivity corresponds to the time when wildtype engrailed protein localizes into segmentally reiterated stripes and represents only a small portion of the normal period of engrailed gene expression.

scholarly journals XACT-seq comprehensively defines the promoter-position and promoter-sequence determinants for initial-transcription pausing

2019 ◽  
Author(s):  
Jared T. Winkelman ◽  
Chirangini Pukhrambam ◽  
Irina O. Vvedenskaya ◽  
Yuanchao Zhang ◽  
Deanne M. Taylor ◽  
...  
Keyword(s):  
Active Center ◽  
Transcription Elongation ◽  
Consensus Sequence ◽  
Promoter Sequence ◽  
Sequence Element ◽  
Promoter Sequences ◽  
Nucleotide Addition ◽  
Sequence Elements ◽  
Nucleotide Resolution

AbstractPausing by RNA polymerase (RNAP) during transcription elongation, in which a translocating RNAP uses a “stepping” mechanism, has been studied extensively, but pausing by RNAP during initial transcription, in which a promoter-anchored RNAP uses a “scrunching” mechanism, has not. We report a method that directly defines RNAP-active-center position relative to DNA in vivo with single-nucleotide resolution (XACT-seq; crosslink-between-active-center-and-template sequencing). We apply this method to detect and quantify pausing in initial transcription at 411 (∼4,000,000) promoter sequences in vivo, in Escherichia coli. The results show initial-transcription pausing can occur in each nucleotide addition during initial transcription, particularly the first 4-5 nucleotide additions. The results further show initial-transcription pausing occurs at sequences that resemble the consensus sequence element for transcription-elongation pausing. Our findings define the positional and sequence determinants for initial-transcription pausing and establish initial-transcription pausing is hard-coded by sequence elements similar to those for transcription-elongation pausing.

Heat shock-induced interactions of heat shock transcription factor and the human hsp70 promoter examined by in vivo footprinting

1991 ◽  
Vol 11 (1) ◽  
pp. 586-592
Author(s):  
K Abravaya ◽  
B Phillips ◽  
R I Morimoto
Keyword(s):  
Transcription Factor ◽  
Heat Shock ◽  
Consensus Sequence ◽  
Regulatory Elements ◽  
Heat Shock Transcription Factor ◽  
Hsp70 Promoter ◽  
Heat Shock Element ◽  
In Vivo Footprinting ◽  
Human Hsp70

Genomic footprinting of the human hsp70 promoter reveals that heat shock induces a rapid binding of a factor, presumably heat shock transcription factor, to a region encompassing five contiguous NGAAN sequences, three perfect and two imperfect matches to the consensus sequence. Arrays of inverted NGAAN sequences have been defined as the heat shock element. No protein is bound to the heat shock element prior to or after recovery from heat shock. Heat shock does not perturb the binding of factors to other regulatory elements in the promoter which contribute to basal expression of the hsp70 gene.

scholarly journals The heat shock consensus sequence is not sufficient for hsp70 gene expression in Drosophila melanogaster.

1985 ◽  
Vol 5 (1) ◽  
pp. 197-203 ◽  
Author(s):  
J Amin ◽  
R Mestril ◽  
R Lawson ◽  
H Klapper ◽  
R Voellmy
Keyword(s):  
Drosophila Melanogaster ◽  
Heat Shock ◽  
Consensus Sequence ◽  
Gene Promoter ◽  
Cell Types ◽  
Hsp70 Gene ◽  
Base Pairs ◽  
Hybrid Gene ◽  
Induced Expression ◽  
African Green Monkey Kidney

A hybrid gene in which the expression of an Escherichia coli beta-galactosidase gene was placed under the control of a Drosophila melanogaster 70,000-dalton heat shock protein (hsp70) gene promoter was constructed. Mutant derivatives of this hybrid gene which contained promoter sequences of different lengths were prepared, and their heat-induced expression was examined in D. melanogaster and COS-1 (African green monkey kidney) cells. Mutants with 5' nontranscribed sequences of at least 90 and up to 1,140 base pairs were expressed strongly in both cell types. Mutants with shorter 5' extensions (of at least 63 base pairs) were transcribed and translated efficiently in COS-1 but not at all in D. melanogaster cells. Thus, in contrast to the situation in COS-1 cells, the previously defined heat shock consensus sequence which is located between nucleotides 62 and 48 of the hsp70 gene 5' nontranscribed DNA segment is not sufficient for the expression of the D. melanogaster gene in homologous cells. A second consensus-like element 69 to 85 nucleotides upstream from the cap site is postulated to be also involved in the heat-induced expression of the hsp70 gene in D. melanogaster cells.

scholarly journals High-resolution mapping of DNase I-hypersensitive sites of Drosophila heat shock genes in Drosophila melanogaster and Saccharomyces cerevisiae.

1984 ◽  
Vol 4 (9) ◽  
pp. 1853-1863 ◽  
Author(s):  
N Costlow ◽  
J T Lis
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Drosophila Melanogaster ◽  
Heat Shock ◽  
High Resolution ◽  
Consensus Sequence ◽  
Dnase I ◽  
Heat Shock Genes ◽  
Base Pairs ◽  
Dnase I Hypersensitive Sites ◽  
Hypersensitive Sites

High-resolution analysis of the chromatin structure of the promoter regions of five Drosophila heat shock genes showed a similar location for the hypersensitive sequences relative to the start of transcription. For each of the five genes examined--those coding for hsp27, hsp26, hsp23, hsp70, and hsp83--the DNase I-hypersensitive sites in Drosophila melanogaster nuclei mapped to two regions upstream of the coding region. These sites occurred on the average, 115 and 17 base pairs upstream from the start of transcription of the five heat shock genes examined. This latter site corresponded to sequences at or near the TATA consensus sequence. Sites even further upstream of the hsp27, hsp26, and hsp83 genes were also evident. Additionally, for the two genes examined--hsp70 and hsp83--the DNase I-hypersensitive sites were preserved, at least within this level of resolution (+/- 10 base pairs), when the Drosophila genes were integrated into the Saccharomyces cerevisiae genome. This result indicates that the signals responsible for generating these hypersensitive sites are inherent in the DNA sequences and, in this case, are not highly species specific.

High-resolution mapping of DNase I-hypersensitive sites of Drosophila heat shock genes in Drosophila melanogaster and Saccharomyces cerevisiae

1984 ◽  
Vol 4 (9) ◽  
pp. 1853-1863
Author(s):  
N Costlow ◽  
J T Lis
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Drosophila Melanogaster ◽  
Heat Shock ◽  
High Resolution ◽  
Consensus Sequence ◽  
Dnase I ◽  
Heat Shock Genes ◽  
Base Pairs ◽  
Dnase I Hypersensitive Sites ◽  
Hypersensitive Sites

High-resolution analysis of the chromatin structure of the promoter regions of five Drosophila heat shock genes showed a similar location for the hypersensitive sequences relative to the start of transcription. For each of the five genes examined--those coding for hsp27, hsp26, hsp23, hsp70, and hsp83--the DNase I-hypersensitive sites in Drosophila melanogaster nuclei mapped to two regions upstream of the coding region. These sites occurred on the average, 115 and 17 base pairs upstream from the start of transcription of the five heat shock genes examined. This latter site corresponded to sequences at or near the TATA consensus sequence. Sites even further upstream of the hsp27, hsp26, and hsp83 genes were also evident. Additionally, for the two genes examined--hsp70 and hsp83--the DNase I-hypersensitive sites were preserved, at least within this level of resolution (+/- 10 base pairs), when the Drosophila genes were integrated into the Saccharomyces cerevisiae genome. This result indicates that the signals responsible for generating these hypersensitive sites are inherent in the DNA sequences and, in this case, are not highly species specific.

The heat shock consensus sequence is not sufficient for hsp70 gene expression in Drosophila melanogaster

1985 ◽  
Vol 5 (1) ◽  
pp. 197-203
Author(s):  
J Amin ◽  
R Mestril ◽  
R Lawson ◽  
H Klapper ◽  
R Voellmy
Keyword(s):  
Drosophila Melanogaster ◽  
Heat Shock ◽  
Consensus Sequence ◽  
Gene Promoter ◽  
Cell Types ◽  
Hsp70 Gene ◽  
Base Pairs ◽  
Hybrid Gene ◽  
Induced Expression ◽  
African Green Monkey Kidney

A hybrid gene in which the expression of an Escherichia coli beta-galactosidase gene was placed under the control of a Drosophila melanogaster 70,000-dalton heat shock protein (hsp70) gene promoter was constructed. Mutant derivatives of this hybrid gene which contained promoter sequences of different lengths were prepared, and their heat-induced expression was examined in D. melanogaster and COS-1 (African green monkey kidney) cells. Mutants with 5' nontranscribed sequences of at least 90 and up to 1,140 base pairs were expressed strongly in both cell types. Mutants with shorter 5' extensions (of at least 63 base pairs) were transcribed and translated efficiently in COS-1 but not at all in D. melanogaster cells. Thus, in contrast to the situation in COS-1 cells, the previously defined heat shock consensus sequence which is located between nucleotides 62 and 48 of the hsp70 gene 5' nontranscribed DNA segment is not sufficient for the expression of the D. melanogaster gene in homologous cells. A second consensus-like element 69 to 85 nucleotides upstream from the cap site is postulated to be also involved in the heat-induced expression of the hsp70 gene in D. melanogaster cells.

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