Sequences involved in temperature and ecdysterone-induced transcription are located in separate regions of a Drosophila melanogaster heat 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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Upstream sequences required for efficient expression of a soybean heat shock gene.

Molecular and Cellular Biology ◽

10.1128/mcb.6.2.559 ◽

1986 ◽

Vol 6 (2) ◽

pp. 559-565 ◽

Cited By ~ 30

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.

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The ovarian, ecdysterone, and heat-shock-responsive promoters of the Drosophila melanogaster hsp27 gene react very differently to perturbations of DNA sequence

Molecular and Cellular Biology ◽

10.1128/mcb.7.3.973-981.1987 ◽

1987 ◽

Vol 7 (3) ◽

pp. 973-981

Author(s):

E P Hoffman ◽

S L Gerring ◽

V G Corces

Keyword(s):

Drosophila Melanogaster ◽

Heat Shock ◽

Dna Sequence ◽

Germ Line ◽

P Element ◽

Position Effects ◽

Consensus Sequences ◽

Basal Expression ◽

Upstream Promoter ◽

Germ Line Transformation

The effect of various types of DNA sequence alterations on the activity of the ovarian, ecdysterone, and heat-shock-responsive promoters of the Drosophila melanogaster hsp27 gene was studied by P element-mediated germ line transformation. Regions of DNA required for proper expression of the gene under these different conditions were identified. Wild-type levels of transcription during oogenesis are dependent on two elements respectively located within a 64-base-pair (bp) fragment in the transcribed untranslated region and between -227 and -958 bp upstream of the transcription start site. This ovarian expression is particularly sensitive to both chromosomal position effects and an increased distance between the distal upstream promoter element and the TATAA homology. The ecdysterone-mediated expression during metamorphosis is dependent on a 145-bp domain including the TATAA box and additional upstream sequences that augment transcription by two- to five-fold. Finally, sequences necessary for heat shock expression are located much further upstream from hsp27 than those previously found for hsp70, although basal expression was correlated with the presence of more proximal heat shock consensus sequences.

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

Molecular and Cellular Biology ◽

10.1128/mcb.6.2.559-565.1986 ◽

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.

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Hsp28stl: A P-Element Insertion Mutation That Alters the Expression of a Heat Shock Gene in Drosophila melanogaster

Genetics ◽

10.1093/genetics/115.2.333 ◽

1987 ◽

Vol 115 (2) ◽

pp. 333-340

Author(s):

Joel C Eissenberg ◽

Sarah C R Elgin

Keyword(s):

Drosophila Melanogaster ◽

Heat Shock ◽

P Element ◽

Developmental Expression ◽

Heat Shock Gene ◽

Insertion Mutation ◽

Target Sequence ◽

Element Insertion ◽

Large Heat ◽

Shock Gene

ABSTRACT We have identified and cloned a mutant allele of the small heat shock gene Hsp28 of Drosophila melanogaster. This allele, which we have called Hsp28stl, produces small amounts of a single aberrantly large, heat-inducible transcript in heat-shocked flies, while a normal-sized Hsp28 transcript is present only in fertile females. No Hsp28 transcript at all is detected in mutant prepupae, a stage when wild-type flies show high levels of Hsp28 RNA. We have cloned the Hsp28stl allele, and have found that a 1.3-kb defective P-element is present 5' to Hsp28 in the mutant line. The site of P-element insertion lies between the Hsp28 "TATA box" sequence and the Hsp28 RNA cap site; in contrast to previously described P-element insertions, the element at Hsp28stl is flanked by a two base pair duplication of the insertional target sequence. The results suggest that this insert may separate elements regulating heat-inducible and developmental expression of Hsp28, leading to the different patterns of transcription observed.

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The ovarian, ecdysterone, and heat-shock-responsive promoters of the Drosophila melanogaster hsp27 gene react very differently to perturbations of DNA sequence.

Molecular and Cellular Biology ◽

10.1128/mcb.7.3.973 ◽

1987 ◽

Vol 7 (3) ◽

pp. 973-981 ◽

Cited By ~ 25

Author(s):

E P Hoffman ◽

S L Gerring ◽

V G Corces

Keyword(s):

Drosophila Melanogaster ◽

Heat Shock ◽

Dna Sequence ◽

Germ Line ◽

P Element ◽

Position Effects ◽

Consensus Sequences ◽

Basal Expression ◽

Upstream Promoter ◽

Germ Line Transformation

The effect of various types of DNA sequence alterations on the activity of the ovarian, ecdysterone, and heat-shock-responsive promoters of the Drosophila melanogaster hsp27 gene was studied by P element-mediated germ line transformation. Regions of DNA required for proper expression of the gene under these different conditions were identified. Wild-type levels of transcription during oogenesis are dependent on two elements respectively located within a 64-base-pair (bp) fragment in the transcribed untranslated region and between -227 and -958 bp upstream of the transcription start site. This ovarian expression is particularly sensitive to both chromosomal position effects and an increased distance between the distal upstream promoter element and the TATAA homology. The ecdysterone-mediated expression during metamorphosis is dependent on a 145-bp domain including the TATAA box and additional upstream sequences that augment transcription by two- to five-fold. Finally, sequences necessary for heat shock expression are located much further upstream from hsp27 than those previously found for hsp70, although basal expression was correlated with the presence of more proximal heat shock consensus sequences.

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Ecdysteroid-regulated heat-shock gene expression during Drosophila melanogaster development

Developmental Biology ◽

10.1016/0012-1606(86)90263-0 ◽

1986 ◽

Vol 115 (2) ◽

pp. 434-438 ◽

Cited By ~ 38

Author(s):

Steven R. Thomas ◽

Judith A. Lengyel

Keyword(s):

Gene Expression ◽

Drosophila Melanogaster ◽

Heat Shock ◽

Heat Shock Gene ◽

Shock Gene ◽

Heat Shock Gene Expression

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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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