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

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.

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

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.

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.

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.

Coordinate changes in heat shock element-binding activity and HSP70 gene transcription rates in human cells

1988 ◽  
Vol 8 (11) ◽  
pp. 4736-4744
Author(s):  
D D Mosser ◽  
N G Theodorakis ◽  
R I Morimoto
Keyword(s):  
Protein Synthesis ◽  
Amino Acid ◽  
Heat Shock ◽  
Gene Transcription ◽  
Elevated Temperatures ◽  
Binding Activity ◽  
Heat Shock Gene ◽  
Hsp70 Gene ◽  
Heat Shock Element ◽  
Amino Acid Analogs

Activation of human heat shock gene transcription by heat shock, heavy metal ions, and amino acid analogs required the heat shock element (HSE) in the HSP70 promoter. Both heat shock- and metal ion-induced HSP70 gene transcription occurred independently of protein synthesis, whereas induction by amino acid analogs required protein synthesis. We identified a HSE-binding activity from control cells which was easily distinguished by a gel mobility shift assay from the stress-induced HSE-binding activity which appeared following heat shock or chemically induced stress. The kinetics of HSP70 gene transcription paralleled the rapid appearance of stress-induced HSE-binding activity. During recovery from heat shock, both the rate of HSP70 gene transcription and stress-induced HSE-binding activity levels declined and the control HSE-binding activity reappeared. The DNA contacts of the control and stress-induced HSE-binding activities deduced by methylation interference were similar but not identical. While stable complexes with HSE were formed with extracts from both control and stressed cells in vitro at 25 degrees C, only the stress-induced complex was detected when binding reactions were performed at elevated temperatures.

scholarly journals Expression of a constitutively active mutant of heat shock factor 1 under the control of testis-specific hst70 gene promoter in transgenic mice induces degeneration of seminiferous epithelium.

2003 ◽  
Vol 50 (2) ◽  
pp. 535-541 ◽  
Author(s):  
Wiesława Widłak ◽  
Konrad Benedyk ◽  
Natallia Vydra ◽  
Magdalena Głowala ◽  
Dorota Scieglińska ◽  
...  
Keyword(s):  
Heat Shock ◽  
Transgenic Mice ◽  
Target Genes ◽  
Gene Promoter ◽  
Basic Mechanism ◽  
Seminiferous Epithelium ◽  
High Rate ◽  
Heat Shock Factor 1 ◽  
Heat Shock Element ◽  
Constitutively Active

Heat shock activates in somatic cells a set of genes encoding heat shock proteins which function as molecular chaperones. The basic mechanism by which these genes are activated is the interaction of the specific transcription factor HSF1 with a regulatory DNA sequence called heat shock element (HSE). In higher eukaryotes HSF1 is present in unstressed cells as inactive monomers which, in response to cellular stress, aggregate into transcriptionally competent homotrimers. In the present paper we showed that the expression of a transgene encoding mutated constitutively active HSF1 placed under the control of a spermatocyte-specific promoter derived from the hst70 gene severely affects spermatogenesis. We found the testes of transgenic mice to be significantly smaller than those of wild-type males and histological analysis showed massive degeneration of the seminiferous epithelium. The lumen of tubules was devoid of spermatids and spermatozoa and using the TUNEL method we demonstrated a high rate of spermatocyte apoptosis. The molecular mechanism by which constitutively active HSF1 arrests spermatogenesis is not known so far. One can assume that HSF1 can either induce or repress so far unknown target genes involved in germ cell apoptosis.

scholarly journals Identification and sequence analysis of a new member of the mouse HSP70 gene family and characterization of its unique cellular and developmental pattern of expression in the male germ line.

1988 ◽  
Vol 8 (7) ◽  
pp. 2925-2932 ◽  
Author(s):  
Z F Zakeri ◽  
D J Wolgemuth ◽  
C R Hunt
Keyword(s):  
Heat Shock ◽  
Gene Family ◽  
Developmental Stages ◽  
Germ Line ◽  
Cell Lineage ◽  
Hsp70 Gene ◽  
Coding Region ◽  
Male Germ Line ◽  
Sequence Organization

A unique member of the mouse HSP70 gene family has been isolated and characterized with respect to its DNA sequence organization and expression. The gene contains extensive similarity to a heat shock-inducible HSP70 gene within the coding region but diverges in both 3' and 5' nontranslated regions. The gene does not yield transcripts in response to heat shock in mouse L cells. Rather, the gene appears to be activated uniquely in the male germ line. Analysis of RNA from different developmental stages and from enriched populations of spermatogenic cells revealed that this gene is expressed during the prophase stage of meiosis. A transcript different in size from the major heat-inducible mouse transcripts is most abundant in meiotic prophase spermatocytes and decreases in abundance in postmeiotic stages of spermatogenesis. This pattern of expression is distinct from that observed for another member of this gene family, which was previously shown to be expressed abundantly in postmeiotic germ cells. These observations suggest that specific HSP70 gene family members play distinct roles in the differentiation of the germ cell lineage in mammals.

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

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.

Dietary components modulate yolk protein gene transcription in Drosophila melanogaster

Development ◽  
1988 ◽  
Vol 103 (1) ◽  
pp. 119-128 ◽  
Author(s):  
M. Bownes ◽  
A. Scott ◽  
A. Shirras
Keyword(s):  
Protein Synthesis ◽  
Drosophila Melanogaster ◽  
Juvenile Hormone ◽  
Gene Transcription ◽  
Protein Gene ◽  
Yolk Protein ◽  
Complete Medium ◽  
General Effect ◽  
Total Recovery ◽  
Insect Hormones

The three yolk proteins of Drosophila melanogaster begin to be synthesized at eclosion. Transcription of the genes is regulated by the genes tra, tra-2 and dsx and also by the insect hormones, juvenile hormone and 20-hydroxyecdysone. We show that there is yet another level of control which is dependent upon feeding. Females that are starved from eclosion show a basal level of yolk protein gene transcription, which is rapidly increased when a complete diet is supplied. We show that the effect is not due to incorrect development of the fat body and is unlikely to be solely due to a general effect on protein synthesis. Later in development, cessation of feeding leads to selective inhibition of yolk protein synthesis and hence egg production. The effects of starvation can be partially overcome by 20-hydroxyecdysone, juvenile hormone, casein, amino acid mix or sucrose, but only a complete medium or live yeast brings about total recovery. Using yp1-Adh fusions (fusions of the promoter region of yp1 to the structural gene for Adh), the DNA sequence required for this diet-enhanced transcription has been located within an 890 bp fragment upstream of the yp1 gene. The insect hormones do not operate on this same DNA fragment.

A Genetically Marked I Element in Drosophila melanogaster Can Be Mobilized When ORF2 Is Provided in trans

Genetics ◽  
1998 ◽  
Vol 148 (1) ◽  
pp. 267-275
Author(s):  
Isabelle Busseau ◽  
Sophie Malinsky ◽  
Maria Balakireva ◽  
Marie-Christine Chaboissier ◽  
Danielle Teninges ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Heat Shock ◽  
Reverse Transcriptase ◽  
Germ Line ◽  
Ltr Retrotransposons ◽  
Low Frequencies ◽  
High Frequencies ◽  
In Trans ◽  
Very High Frequencies ◽  
Very High

Abstract I factors in Drosophila melanogaster are non-LTR retrotransposons similar to mammalian LINEs. They transpose at very high frequencies in the germ line of SF females resulting from crosses between reactive females, devoid of active I factors, and inducer males, containing active I factors. The vermilion marked IviP2 element was designed to allow easy phenotypical screening for retrotransposition events. It is deleted in ORF2 and therefore cannot produce reverse transcriptase. IviP2 can be mobilized at very low frequencies by actively transposing I factors in the germ line of SF females. This paper shows that IviP2 can be mobilized more efficiently in the germ line of strongly reactive females in the absence of active I factors, when it is trans-complemented by the product of ORF2 synthesized from the hsp70 heat-shock promoter. This represents a promising step toward the use of marked I elements to study retrotransposition and as tools for mutagenesis.

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

1990 ◽  
Vol 10 (6) ◽  
pp. 3232-3238 ◽  
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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