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

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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Sex-specific control of Drosophila melanogaster yolk protein 1 gene expression is limited to transcription

Molecular and Cellular Biology ◽

10.1128/mcb.8.11.4756-4764.1988 ◽

1988 ◽

Vol 8 (11) ◽

pp. 4756-4764

Author(s):

K W Kraus ◽

Y H Lee ◽

J T Lis ◽

M F Wolfner

Keyword(s):

Drosophila Melanogaster ◽

Heat Shock ◽

Gene Transcription ◽

Rna Splicing ◽

Posttranscriptional Regulation ◽

Target Genes ◽

Germ Line ◽

Gene Promoter ◽

Yolk Protein ◽

Hsp70 Gene

The sex of Drosophila melanogaster is determined by a hierarchy of genes. The ultimate targets of this regulatory hierarchy are the genes encoding terminal differentiation products of one sex. For one of the best-characterized target genes, that encoding female-specific yolk protein 1 (YP1), sex-specific transcriptional controls have been clearly demonstrated. In addition, sex-specific posttranscriptional controls were suggested from experiments in which YP1 RNA was induced in males with hormones. To determine whether males can efficiently process and translate a transcript which is normally found only in females, we used a non-sex-specific promoter, the hsp70 gene promoter, to drive YP1 gene transcription in germ line transformed males. The efficiency of expression of the YP1 gene at levels of RNA splicing, translation, and protein secretion in these males was compared with that in wild-type females. These experiments show that there are no sex-specific posttranscriptional controls operating to limit the production of secreted YP1 in males. Promoters containing different numbers of heat shock elements were tested for their ability to drive YP1 gene transcription in males. These results show that incompatibility between the hsp70 gene heat shock elements and the YP1 gene promoter can be overcome by increasing the amount of hsp70 gene sequence up or downstream of the TATA box. In the course of this study, two vectors useful for placing genes under heat shock regulation were constructed. One of these vectors is designed so that the heat-induced transcript produced is the "authentic" primary transcript; it should be useful for studies of posttranscriptional regulation.

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

Molecular and Cellular Biology ◽

10.1128/mcb.4.9.1853 ◽

1984 ◽

Vol 4 (9) ◽

pp. 1853-1863 ◽

Cited By ~ 36

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.

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Sex-specific control of Drosophila melanogaster yolk protein 1 gene expression is limited to transcription.

Molecular and Cellular Biology ◽

10.1128/mcb.8.11.4756 ◽

1988 ◽

Vol 8 (11) ◽

pp. 4756-4764 ◽

Cited By ~ 10

Author(s):

K W Kraus ◽

Y H Lee ◽

J T Lis ◽

M F Wolfner

Keyword(s):

Drosophila Melanogaster ◽

Heat Shock ◽

Gene Transcription ◽

Rna Splicing ◽

Posttranscriptional Regulation ◽

Target Genes ◽

Germ Line ◽

Gene Promoter ◽

Yolk Protein ◽

Hsp70 Gene

The sex of Drosophila melanogaster is determined by a hierarchy of genes. The ultimate targets of this regulatory hierarchy are the genes encoding terminal differentiation products of one sex. For one of the best-characterized target genes, that encoding female-specific yolk protein 1 (YP1), sex-specific transcriptional controls have been clearly demonstrated. In addition, sex-specific posttranscriptional controls were suggested from experiments in which YP1 RNA was induced in males with hormones. To determine whether males can efficiently process and translate a transcript which is normally found only in females, we used a non-sex-specific promoter, the hsp70 gene promoter, to drive YP1 gene transcription in germ line transformed males. The efficiency of expression of the YP1 gene at levels of RNA splicing, translation, and protein secretion in these males was compared with that in wild-type females. These experiments show that there are no sex-specific posttranscriptional controls operating to limit the production of secreted YP1 in males. Promoters containing different numbers of heat shock elements were tested for their ability to drive YP1 gene transcription in males. These results show that incompatibility between the hsp70 gene heat shock elements and the YP1 gene promoter can be overcome by increasing the amount of hsp70 gene sequence up or downstream of the TATA box. In the course of this study, two vectors useful for placing genes under heat shock regulation were constructed. One of these vectors is designed so that the heat-induced transcript produced is the "authentic" primary transcript; it should be useful for studies of posttranscriptional regulation.

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

Molecular and Cellular Biology ◽

10.1128/mcb.4.9.1853-1863.1984 ◽

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.

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Two transcriptional activators, CCAAT-box-binding transcription factor and heat shock transcription factor, interact with a human hsp70 gene promoter

Molecular and Cellular Biology ◽

10.1128/mcb.7.3.1129-1138.1987 ◽

1987 ◽

Vol 7 (3) ◽

pp. 1129-1138

Author(s):

W D Morgan ◽

G T Williams ◽

R I Morimoto ◽

J Greene ◽

R E Kingston ◽

...

Keyword(s):

Transcription Factor ◽

Heat Shock ◽

Gene Promoter ◽

Heat Shock Transcription Factor ◽

Transcriptional Activators ◽

Hsp70 Gene ◽

Nuclear Extracts ◽

Ccaat Box ◽

Human Hsp70

We characterized the activity of a human hsp70 gene promoter by in vitro transcription. Analysis of 5' deletion and substitution mutants in HeLa nuclear extracts showed that the basal activity of the promoter depends primarily on a CCAAT-box sequence located at -65. A protein factor, CCAAT-box-binding transcription factor (CTF), was isolated from HeLa nuclear extracts and shown to be responsible for stimulation of transcription in a reconstituted in vitro system. DNase I footprinting revealed that CTF interacts with two CCAAT-box elements located at -65 and -147 of the human hsp70 promoter. An additional binding activity, heat shock transcription factor (HSTF), which interacted with the heat shock element, was also identified in HeLa extract fractions. This demonstrates that the promoter of this human hsp70 gene interacts with at least two positive transcriptional activators, CTF, which is required for CCAAT-box-dependent transcription as in other promoters such as those of globin and herpes simplex virus thymidine kinase genes, and HSTF, which is involved in heat inducibility.

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Organization of the Drosophila melanogaster hsp70 heat shock regulation unit

Molecular and Cellular Biology ◽

10.1128/mcb.7.3.1055-1062.1987 ◽

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.

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Molecular and developmental characterization of the heat shock cognate 4 gene of Drosophila melanogaster.

Molecular and Cellular Biology ◽

10.1128/mcb.10.6.3232 ◽

1990 ◽

Vol 10 (6) ◽

pp. 3232-3238 ◽

Cited By ~ 48

Author(s):

L A Perkins ◽

J S Doctor ◽

K Zhang ◽

L Stinson ◽

N Perrimon ◽

...

Keyword(s):

Drosophila Melanogaster ◽

Heat Shock ◽

Gene Family ◽

Rapid Growth ◽

Developmental Expression ◽

Hsp70 Gene ◽

Abundant Protein ◽

Cognate Gene ◽

Gene 4

The Drosophila heat shock cognate gene 4 (hsc4), a member of the hsp70 gene family, encodes an abundant protein, hsc70, that is more similar to the constitutively expressed human protein than the Drosophila heat-inducible hsp70. Developmental expression revealed that hsc4 transcripts are enriched in cells active in endocytosis and those undergoing rapid growth and changes in shape.

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Repression of hsp70 heat shock gene transcription by the suppressor of hairy-wing protein of Drosophila melanogaster.

Molecular and Cellular Biology ◽

10.1128/mcb.11.4.1894 ◽

1991 ◽

Vol 11 (4) ◽

pp. 1894-1900 ◽

Cited By ~ 91

Author(s):

C Holdridge ◽

D Dorsett

Keyword(s):

Drosophila Melanogaster ◽

Heat Shock ◽

Protein Binding ◽

Protein Interactions ◽

Binding Sites ◽

Zinc Finger Protein ◽

Gene Promoter ◽

Heat Shock Gene ◽

Protein Binding Sites ◽

Shock Gene

The suppressor of hairy-wing [su(Hw)] locus of Drosophila melanogaster encodes a zinc finger protein that binds a repeated motif in the gypsy retroposon. Mutations of su(Hw) suppress the phenotypes associated with mutations caused by gypsy insertions. To examine the mechanisms by which su(Hw) alters gene expression, a fragment of gypsy containing multiple su(Hw) protein-binding sites was inserted into various locations in the well-characterized Drosophila hsp70 heat shock gene promoter. We found no evidence for activation of basal hsp70 transcription by su(Hw) protein in cultured Drosophila cells but observed that it can repress heat shock-induced transcription. Repression occurred only when su(Hw) protein-binding sites were positioned between binding sites for proteins required for heat shock transcription. We propose that su(Hw) protein interferes nonspecifically with protein-protein interactions required for heat shock transcription, perhaps sterically, or by altering the ability of DNA to bend or twist.

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

Molecular and Cellular Biology ◽

10.1128/mcb.6.2.663 ◽

1986 ◽

Vol 6 (2) ◽

pp. 663-673 ◽

Cited By ~ 32

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.

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