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.

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 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.

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

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.

In vivo interactions of RNA polymerase II with genes of Drosophila melanogaster

1985 ◽  
Vol 5 (8) ◽  
pp. 2009-2018
Author(s):  
D S Gilmour ◽  
J T Lis
Keyword(s):  
Gene Expression ◽  
Drosophila Melanogaster ◽  
Heat Shock ◽  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Protein Characterization ◽  
Heat Shock Gene ◽  
Transcriptional Level ◽  
Intact Cells

We describe a method for examining the in vivo distribution of a protein on specific eucaryotic DNA sequences. In this method, proteins are cross-linked to DNA in intact cells, and the protein-DNA adducts are isolated by immunoprecipitation with antiserum against the protein. Characterization of the DNA cross-linked to the precipitated protein identifies the sequences with which the protein is associated in vivo. Here, we applied these methods to detect RNA polymerase II-DNA interactions in heat-shocked and untreated Drosophila melanogaster Schneider line 2 cells. The level of RNA polymerase II associated with several heat shock genes increased dramatically in response to heat shock, whereas the level associated with the copia genes decreased, indicating that both induction of heat shock gene expression and repression of the copia gene expression by heat shock occur at the transcriptional level. Low levels of RNA polymerase II were present on DNA outside of the transcription units, and for at least two genes, hsp83 and hsp26, RNA polymerase II initiated binding near the transcription start site. Moreover, for hsp70, the density of RNA polymerase II on sequences downstream of the polyadenylate addition site was much lower than that observed on the gene internal sequences. Examination of the amount of specific restriction fragments cross-linked to RNA polymerase II provides a means of detecting RNA polymerase II on individual members of multigene families. This analysis shows that RNA polymerase II is associated with only one of the two cytoplasmic actin genes.

Chromatin structure of a P-element-transduced hsp-28 gene in Drosophila melanogaster

1986 ◽  
Vol 6 (11) ◽  
pp. 4126-4129
Author(s):  
J C Eissenberg ◽  
S C Elgin
Keyword(s):  
Gene Expression ◽  
Drosophila Melanogaster ◽  
Chromatin Structure ◽  
Dnase I ◽  
P Element ◽  
Direct Selection ◽  
Wild Type ◽  
Dnase I Hypersensitive Sites ◽  
Hypersensitive Sites ◽  
Selection For

The Drosophila hsp-28 gene was heat inducible when transduced to novel chromosomal sites even when no direct selection for transduced gene expression was imposed. The pattern of DNase I-hypersensitive sites 5' to the wild type and transduced copy of hsp-28 was similar. In addition, DNase I-hypersensitive sites occurred within the P-element sequences flanking transduced loci.

Genetic and molecular characterization of a variegating hsp70-lacZ fusion gene in the euchromatic 31B region of Drosophila melanogaster

Genome ◽  
2001 ◽  
Vol 44 (4) ◽  
pp. 698-707
Author(s):  
Patrick Morcillo ◽  
Ross J MacIntyre
Keyword(s):  
Drosophila Melanogaster ◽  
Heat Shock ◽  
Fusion Gene ◽  
Position Effect ◽  
P Element ◽  
Lacz Fusion ◽  
Position Effect Variegation ◽  
Salivary Gland Cells ◽  
P Element Transformation

A hsp70–lacZ fusion gene introduced into Drosophila melanogaster at the euchromatic 31B region by P-element transformation displayed a variegated expression with respect to the lacZ fusion protein in the salivary gland cells under heat-shock conditions. The variegation is also reflected by the chromosome puffing pattern. Subsequent transposition of the 31B P element to other euchromatic positions restored wild-type activity, that is, a nonvariegated phenotype. A lower developmental temperature reduced the amount of expression under heat-shock conditions, similar to genes undergoing position-effect variegation (PEV). However, other modifiers of PEV did not affect the expression pattern of the gene. These results show a novel euchromatic tissue-specific variegation that is not associated with classical heterochromatic PEV.Key words: Drosophila, euchromatic position effect, heat shock construct.

scholarly journals Modification of Heat-Shock Gene Expression in Drosophila melanogaster Populations via Transposable Elements

2003 ◽  
Vol 20 (1) ◽  
pp. 135-144 ◽  
Author(s):  
Daniel N. Lerman ◽  
Pawel Michalak ◽  
Amanda B. Helin ◽  
Brian R. Bettencourt ◽  
Martin E. Feder
Keyword(s):  
Gene Expression ◽  
Drosophila Melanogaster ◽  
Heat Shock ◽  
Transposable Elements ◽  
Heat Shock Gene ◽  
Shock Gene ◽  
Heat Shock Gene Expression
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