scholarly journals Multiple, compensatory regulatory elements specify spermatocyte-specific expression of the Drosophila melanogaster hsp26 gene.

1990 ◽  
Vol 10 (1) ◽  
pp. 131-137 ◽  
Author(s):  
R L Glaser ◽  
J T Lis
Keyword(s):  
Drosophila Melanogaster ◽  
Nurse Cell ◽  
Germ Line ◽  
Regulatory Elements ◽  
Heterologous Promoter ◽  
Specific Expression ◽  
Promoter Sequences ◽  
General Response ◽  
Redundant Functions ◽  
Germ Line Transformation

The hsp26 gene of Drosophila melanogaster is expressed in six tissues during development and in a tissue-general response to heat shock. To be able to compare tissue-specific and heat-induced mechanisms of hsp26 expression, we have begun an analysis of the sequences involved in the spermatocyte-specific expression of the hsp26 gene by using germ line transformation. hsp26 mRNA synthesized in the spermatocytes has the same start site as sites previously demonstrated for nurse cell-specific and heat-induced mRNAs. Three regions of the hsp26 gene (nucleotides -351 to -135, -135 to -85, and +11 to +632) were able to stimulate spermatocyte-specific expression when fused with promoter sequences (nucleotides -85 to +11) that alone were insufficient to stimulate expression. These stimulatory regions appear to contain elements that provide redundant functions. While each region was able to stimulate expression independently, the deletion of any one region from a construct was without consequence as long as another compensatory region(s) was still present. There must reside, at a minimum, two independent spermatocyte-specifying elements within the sequences that encompass the three stimulatory regions and the promoter. At least one element is contained within sequences from -351 to -48. This region, in either orientation, can stimulate spermatocyte-specific expression from a heterologous promoter. A second element must reside in sequences from -52 to +632, since these sequences are also sufficient to direct spermatocyte-specific expression.

Multiple, compensatory regulatory elements specify spermatocyte-specific expression of the Drosophila melanogaster hsp26 gene

1990 ◽  
Vol 10 (1) ◽  
pp. 131-137
Author(s):  
R L Glaser ◽  
J T Lis
Keyword(s):  
Drosophila Melanogaster ◽  
Nurse Cell ◽  
Germ Line ◽  
Regulatory Elements ◽  
Heterologous Promoter ◽  
Specific Expression ◽  
Promoter Sequences ◽  
General Response ◽  
Redundant Functions ◽  
Germ Line Transformation

The hsp26 gene of Drosophila melanogaster is expressed in six tissues during development and in a tissue-general response to heat shock. To be able to compare tissue-specific and heat-induced mechanisms of hsp26 expression, we have begun an analysis of the sequences involved in the spermatocyte-specific expression of the hsp26 gene by using germ line transformation. hsp26 mRNA synthesized in the spermatocytes has the same start site as sites previously demonstrated for nurse cell-specific and heat-induced mRNAs. Three regions of the hsp26 gene (nucleotides -351 to -135, -135 to -85, and +11 to +632) were able to stimulate spermatocyte-specific expression when fused with promoter sequences (nucleotides -85 to +11) that alone were insufficient to stimulate expression. These stimulatory regions appear to contain elements that provide redundant functions. While each region was able to stimulate expression independently, the deletion of any one region from a construct was without consequence as long as another compensatory region(s) was still present. There must reside, at a minimum, two independent spermatocyte-specifying elements within the sequences that encompass the three stimulatory regions and the promoter. At least one element is contained within sequences from -351 to -48. This region, in either orientation, can stimulate spermatocyte-specific expression from a heterologous promoter. A second element must reside in sequences from -52 to +632, since these sequences are also sufficient to direct spermatocyte-specific expression.

Transformation mapping of the regulatory elements of the ecdysone-inducible P1 gene of Drosophila melanogaster

1991 ◽  
Vol 11 (5) ◽  
pp. 2913-2917
Author(s):  
F Maschat ◽  
M L Dubertret ◽  
J A Lepesant
Keyword(s):  
Drosophila Melanogaster ◽  
Hormonal Regulation ◽  
Germ Line ◽  
Regulatory Elements ◽  
Fat Bodies ◽  
Germ Line Transformation ◽  
Third Instar Larvae

The transcription of the P1 gene is induced by 20-hydroxyecdysone in fat bodies of third-instar larvae. Germ line transformation showed that sequences between -138 to +276 contain elements required for a qualitatively correct developmental and hormonal regulation of P1 transcription. Sequences from -138 to -68 are essential for this expression.

scholarly journals Transformation mapping of the regulatory elements of the ecdysone-inducible P1 gene of Drosophila melanogaster.

1991 ◽  
Vol 11 (5) ◽  
pp. 2913-2917 ◽  
Author(s):  
F Maschat ◽  
M L Dubertret ◽  
J A Lepesant
Keyword(s):  
Drosophila Melanogaster ◽  
Hormonal Regulation ◽  
Germ Line ◽  
Regulatory Elements ◽  
Fat Bodies ◽  
Germ Line Transformation ◽  
Third Instar Larvae

The transcription of the P1 gene is induced by 20-hydroxyecdysone in fat bodies of third-instar larvae. Germ line transformation showed that sequences between -138 to +276 contain elements required for a qualitatively correct developmental and hormonal regulation of P1 transcription. Sequences from -138 to -68 are essential for this expression.

scholarly journals cis-acting sequences required for expression of the divergently transcribed Drosophila melanogaster Sgs-7 and Sgs-8 glue protein genes

1991 ◽  
Vol 11 (6) ◽  
pp. 2971-2979
Author(s):  
A Hofmann ◽  
M D Garfinkel ◽  
E M Meyerowitz
Keyword(s):  
Germ Line ◽  
Fusion Gene ◽  
Regulatory Elements ◽  
Sequence Motifs ◽  
Specific Expression ◽  
Cis Acting ◽  
Gene Regulatory Elements ◽  
Glue Protein ◽  
Specific Expression Pattern ◽  
Germ Line Transformation

The Sgs-7 and Sgs-8 glue genes at 68C are divergently transcribed and are separated by 475 bp. Fusion genes with Adh or lacZ coding sequences were constructed, and the expression of these genes, with different amounts of upstream sequences present, was tested by a transient expression procedure and by germ line transformation. A cis-acting element for both genes is located asymmetrically in the intergenic region between -211 and -43 bp relative to Sgs-7. It is required for correct expression of both genes. This element can confer the stage- and tissue-specific expression pattern of glue genes on a heterologous promoter. An 86-bp portion of the element, from -133 to -48 bp relative to Sgs-7, is shown to be capable of enhancing the expression of a truncated and therefore weakly expressed Sgs-3 fusion gene. Recently described common sequence motifs of glue gene regulatory elements (T. Todo, M. Roark, K. Vijay Raghavan, C. A. Mayeda, and E.M. Meyerowitz, Mol. Cell. Biol. 10:5991-6002, 1990) are located within this 86-bp region.

scholarly journals cis-acting sequences required for expression of the divergently transcribed Drosophila melanogaster Sgs-7 and Sgs-8 glue protein genes.

1991 ◽  
Vol 11 (6) ◽  
pp. 2971-2979 ◽  
Author(s):  
A Hofmann ◽  
M D Garfinkel ◽  
E M Meyerowitz
Keyword(s):  
Germ Line ◽  
Fusion Gene ◽  
Regulatory Elements ◽  
Sequence Motifs ◽  
Specific Expression ◽  
Cis Acting ◽  
Gene Regulatory Elements ◽  
Glue Protein ◽  
Specific Expression Pattern ◽  
Germ Line Transformation

The Sgs-7 and Sgs-8 glue genes at 68C are divergently transcribed and are separated by 475 bp. Fusion genes with Adh or lacZ coding sequences were constructed, and the expression of these genes, with different amounts of upstream sequences present, was tested by a transient expression procedure and by germ line transformation. A cis-acting element for both genes is located asymmetrically in the intergenic region between -211 and -43 bp relative to Sgs-7. It is required for correct expression of both genes. This element can confer the stage- and tissue-specific expression pattern of glue genes on a heterologous promoter. An 86-bp portion of the element, from -133 to -48 bp relative to Sgs-7, is shown to be capable of enhancing the expression of a truncated and therefore weakly expressed Sgs-3 fusion gene. Recently described common sequence motifs of glue gene regulatory elements (T. Todo, M. Roark, K. Vijay Raghavan, C. A. Mayeda, and E.M. Meyerowitz, Mol. Cell. Biol. 10:5991-6002, 1990) are located within this 86-bp region.

Delimiting regulatory sequences of the Drosophila melanogaster Ddc gene

1986 ◽  
Vol 6 (12) ◽  
pp. 4548-4557
Author(s):  
J Hirsh ◽  
B A Morgan ◽  
S B Scholnick
Keyword(s):  
Drosophila Melanogaster ◽  
Germ Line ◽  
Regulatory Elements ◽  
P Element ◽  
Developmental Expression ◽  
Upstream Region ◽  
Regulatory Sequences ◽  
Flanking Sequences

We delimited sequences necessary for in vivo expression of the Drosophila melanogaster dopa decarboxylase gene Ddc. The expression of in vitro-altered genes was assayed following germ line integration via P-element vectors. Sequences between -209 and -24 were necessary for normally regulated expression, although genes lacking these sequences could be expressed at 10 to 50% of wild-type levels at specific developmental times. These genes showed components of normal developmental expression, which suggests that they retain some regulatory elements. All Ddc genes lacking the normal immediate 5'-flanking sequences were grossly deficient in larval central nervous system expression. Thus, this upstream region must contain at least one element necessary for this expression. A mutated Ddc gene without a normal TATA boxlike sequence used the normal RNA start points, indicating that this sequences is not required for start point specificity.

scholarly journals Repression and activation of the Drosophila dopa decarboxylase gene in glia.

1992 ◽  
Vol 12 (12) ◽  
pp. 5659-5666 ◽  
Author(s):  
G S Mastick ◽  
S B Scholnick
Keyword(s):  
Germ Line ◽  
Transcriptional Start Site ◽  
Regulatory Region ◽  
Regulatory Elements ◽  
Dopa Decarboxylase ◽  
In Vitro Mutagenesis ◽  
Wild Type ◽  
Cell Activator ◽  
Germ Line Transformation

Glial expression of the Drosophila dopa decarboxylase gene (Ddc) is repressed by a regulatory region located approximately 1 kb upstream of the transcriptional start site. We have used in vitro mutagenesis and germ line transformation to determine which elements within the Ddc promoter mediate repression. Our evidence suggests that the hypodermal cell activator elements IIA and IIB play a major role in the transcriptional regulation of Ddc in glial cells. A variety of mutations demonstrate that element IIA is a strong glial activator element and that element IIB is necessary for glial repression. Although these two regulatory elements are nearly identical in sequence, our data suggest that they are not redundant. Altering the wild-type number and spacing of elements IIA and IIB indicates that the wild-type arrangement of this repeat is critical for repression. We conclude that these key elements of the Ddc promoter regulate both activation and repression in glia.

scholarly journals Seven genes of the Enhancer of split complex of Drosophila melanogaster encode helix-loop-helix proteins.

Genetics ◽  
1992 ◽  
Vol 132 (2) ◽  
pp. 505-518 ◽  
Author(s):  
E Knust ◽  
H Schrons ◽  
F Grawe ◽  
J A Campos-Ortega
Keyword(s):  
Drosophila Melanogaster ◽  
Epidermal Cell ◽  
Germ Line ◽  
Gene Complex ◽  
Helix Loop Helix ◽  
Early Neurogenesis ◽  
Enhancer Of Split ◽  
Germ Line Transformation ◽  
Epidermal Development

Abstract Enhancer of split [E(spl)] is one of the neurogenic loci of Drosophila and, as such, is required for normal segregation of neural and epidermal cell progenitors. Genetic observations indicate that the E(spl) locus is in fact a gene complex comprising a cluster of related genes and that other genes of the region are also required for normal early neurogenesis. Three of the genes of the complex were known to encode helix-loop-helix (HLH) proteins and to be transcribed in nearly identical patterns. Here, we show that four other genes in the vicinity also encode HLH proteins and, during neuroblast segregation, three of them are expressed in the same pattern. We show by germ-line transformation that these three genes are also necessary to allow epidermal development of the neuroectodermal cells.

scholarly journals A DNA segment controlling metal-regulated expression of the Drosophila melanogaster metallothionein gene Mtn

1987 ◽  
Vol 7 (5) ◽  
pp. 1710-1715
Author(s):  
E Otto ◽  
J M Allen ◽  
J E Young ◽  
R D Palmiter ◽  
G Maroni
Keyword(s):  
Drosophila Melanogaster ◽  
Mammalian Cells ◽  
Germ Line ◽  
Initiation Site ◽  
P Element ◽  
Specific Expression ◽  
Metallothionein Gene ◽  
Regulated Expression ◽  
Metallothionein Promoter ◽  
Simplex Virus

Cloned fragments of DNA including the Drosophila melanogaster metallothionein gene Mtn and different amounts of 5' flanking sequences were introduced into flies by P-element-mediated germ line transformation. Comparison of RNA levels in different transformants revealed that metal-regulated and tissue-specific expression of Mtn requires no more than 373 base pairs upstream of the initiation site of transcription. Transformants having an additional, transcribed copy of Mtn could tolerate increased concentrations of cadmium, indicating that Mtn expression is directly related to this phenotype. In separate experiments, these D. melanogaster promoter sequences were fused to the coding sequences of the herpes simplex virus thymidine kinase (TK) gene. After transfection of this fusion into baby hamster kidney cells, increases in TK activity and accumulation of TK RNA were inducible by metals. A series of 5' and 3' deletions showed that D. melanogaster sequences from -130 to -6 were sufficient to confer metal-regulated expression to the TK gene. The function of the D. melanogaster metallothionein promoter in mammalian cells indicates that the mechanism controlling metal regulation is evolutionarily conserved.

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