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G Korge

Publications and source records attributed to G Korge.

25 records · Page 2Linked to original sources

Regulatory sequences of the Sgs-4 gene of Drosophila melanogaster analysed by P element-mediated transformation.

The X chromosomally located allele Sgs-4c for a larval secretion protein of Drosophila melanogaster is normally expressed in female larvae of the strain Oregon R and is hyperexpressed in male larvae exhibiting dosage compensation; the allele Sgs-4d in the strain Samarkand is weakly expressed and is not hyperexpressed in male larvae showing a dosage effect. P element-mediated transformation of upstream DNA sequences from both alleles combined with Sgs-4d coding and downstream sequences was performed to localize sequences which are responsible for the level of gene expression and for hyperexpression of Sgs-4c in male larvae. Our results demonstrate that weak expression and dosage effect are inherited with the upstream region from -1 to -838. This Samarkand fragment differs from the homologous Oregon R region only by a C to T transition at -344 which lies within an assumed binding sequence for the ecdysone receptor complex of dyad base symmetry. Replacing the Samarkand upstream region from -1 to -838 by the Oregon R region restores normal Sgs-4 expression and dosage compensation. Hyperexpression in male larvae displays high sensitivity to position effect and is nearly completely inhibited in one transformed line under heterozygous conditions. The integration of an Sgs-4d transposon into a weak spot of polytene chromosome 2L results in a decrease in gene expression. The GTT- and GT-rich regions at -1.2 and -2.0 kb do not obviously influence Sgs-4 expression but possibly play a role in induction of stage-specific chromosome puffing.

Animals↗

Transformation of salivary gland secretion protein gene Sgs-4 in Drosophila: stage- and tissue-specific regulation, dosage compensation, and position effect.

The Sgs-4 gene of Drosophila melanogaster encodes one of the larval secretion proteins and is active only in salivary glands at the end of larval development. This gene lies in the X chromosome and is controlled by dosage compensation--i.e., the gene is hyperexpressed in males. Therefore, males with one X chromosome produce nearly as much Sgs-4 products as females with two X chromosomes. We used a 4.9-kilobase-pair (kb) DNA fragment containing the Sgs-4d coding region embedded in 2.6 kb of upstream sequences and 1.3 kb of downstream sequences for P-element-mediated transformation of the Sgs-4h underproducer strain Kochi-R. Sgs-4d gene expression was found in all 15 transformed lines analyzed, varying with the site of chromosomal integration. The transposed gene was subject to tissue- and stage-specific regulation. At X-chromosomal sites, the levels of gene expression were similar in both sexes, signifying dosage compensation. At autosomal sites, it was on average 1.5 times higher in males than in females. The results indicate that the transforming DNA fragment contains all sequences necessary for tissue- and stage-specific regulation and for hyperexpression in males.

Age Factors↗

Genetic analysis of the larval secretion gene Sgs-4 and its regulatory chromosome sites in Drosophila melanogaster.

Larval salivary gland secretion from seven wild-type stocks of Drosophila melanogaster was electrophoretically analyzed. Considerable variability occurs in the X-chromosomally coded secretion protein 4, both qualitatively, as expressed by differences in electrophoretic mobilities, and quantitatively as seen by its relative amount in the secretion. Drosophila stocks with "normal" amounts of protein 4 show approximately 80-90% dosage compensation in the males, whereas in two stocks with lower amounts of protein 4 there is no indication of dosage compensation. Genetic analysis showed that the properties of secretion protein 4 and the level of expression of the Sgs-4 gene are controlled by the X-chromosome. Recombination experiments indicate that the stock-specific characteristics of protein 4 are properties of the structural gene Sgs-4 itself or of a chromosome region immediately adjacent to Sgs-4. One recombinant (R+79), manifesting an intermediate level of dosage compensation, indicates that a chromosome segment closely distal to Sg-4 is responsible for the regulation of the gene and for dosage compensation in particular. Accordingly, Sgs-4 must be transcribed from distal to proximal. Its position on the genetic map is 3.6. Two stocks, Hikone-R and Kochi-R, which were originally described as 0-mutants produce very low amounts of a specific secretion protein, 4 h, as revealed by a transvection effect and also by fluorography of overloaded gels.

Animals↗

Direct correlation between a chromosome puff and the synthesis of a larval saliva protein in Drosophila melanogaster.

The structural gene Sgs-4 responsible for larval saliva protein 4 of Drosophila melanogaster was localized, with the help of Notch deficiencies, within the section between bands 3C10 and 3D1 of the X chromosome. In this chromosome section there is, very probably, only one fine band. In the third larval instar chromosome this section is transcriptionally active and forms a puff. When the ecdysone concentration increases, about 5 h before prepupa formation, it becomes inactive.--In section 3C of X chromosomes of third instar larvae of the stock Hikone-R no puff is formed. The saliva of these larvae lacks protein 4. However, female hybrids (H/B and H/O) from Hikone-R crossed with Berlin and Oregon respectively produce a Hikone-specific saliva protein 4h. The synthesis of protein 4h in the hybrids H/B and H/O is ascribed to an activation of the gene Sgs-4 in the Hikone chromosome.--In the saliva of heterozygotes (FM1/H) carrying one inversion chromosome In(1)FM1 and one X chromosome from Hikone, protein 4h could not be detected. In these inversion heterozygotes in 90% of all nuclei the homologues are not paired in 3C, and 3C is puffed only in the FM1 chromosome. This suggests that a precondition for the activation of Hikone gene Sgs-4 in heterozygotes may be intimate homologue pairing.--Intersexes with one of their X chromosomes from Hikone-R and the other from Berlin produce relatively more protein 4h than do diploid H/B females, indicating facilitated transcription as a result of dosage compensation.

Animals↗

Chromosome puff activity and protein synthesis in larval salivary glands of Drosophila melanogaster.

Secretion proteins from larval salivary glands of Drosophila melanogaster were analyzed with acrylamide gel electrophoresis. Four fractions were found; three showed electrophoretic variants in different wild-type stocks. Crossbreeding and cytogenetic techniques were used to localize the genes responsible for the two main fractions: The gene for fraction 3 was found to lie within a segment of the third chromosome which includes section 68C; the gene for fraction 4, Sgs-4, was found to lie within section 3C8-3D1 of the X chromosome (1 - 3-5). The puffs within these sections of the giant chromosomes are active before and during secretion synthesis and become inactive as secretion synthesis ceases. Larvae of one wild-type stock which lack protein fraction 4 do not exhibit any puffing in 3C. The relative amount of protein 4 in the salivary secretion shows a marked dependence on the dosage of the Sgs-4 gene in both duplication and deficiency genotypes. The active site within puff 3C11-12 apparently contains the structural gene for protein 4.

Chromosome Mapping↗