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G Korge

Publications and source records attributed to G Korge.

At least 19 recordsLinked to original sources
PubMed2006

Molecular and immunological characterization of encoding gene and 14-3-3 protein 1 in Fasciola gigantica.

A cDNA encoding Fg14-3-3 protein 1 was cloned by immunoscreening of an adult-stage Fasciola gigantica cDNA library using a rabbit antiserum against tegumental antigens of the parasite. The protein has a deduced amino acid sequence of 252 residues and a calculated molecular weight of 28.7 kDa. It shows sequence identity values between 57.6 and 58.1% to the human 14-3-3 beta, zeta, theta, and eta proteins and is in a phylogenetic cluster with the 14-3-3 protein 1 of Schistosoma spp. Nucleic acid analyses indicate that the Fg14-3-3 protein 1 is encoded by a single copy gene and that this gene is expressed as a transcript of 1250 nucleotides. In adult and 4-week-old parasites the gene's transcriptional and translational products were localized in the gut epithelium, parenchyma, tegument cells, and in the reproductive organs. An antiserum against recombinant Fg14-3-3 protein 1 detected a slightly smaller 14-3-3 protein in the parasite's excretion/secretion material and showed cross-reactivity with 14-3-3 proteins in extracts of other trematodes and mouse. Antibodies against Fg14-3-3 protein were detected in the sera of rabbits as early as 2 weeks after infection with metacercariae of F. gigantica and the antibody titre increased continuously over a 10-week observation period.

14-3-3 Proteins↗
PubMed2001

Innervation of the ring gland of Drosophila melanogaster.

In insects, peptidergic neurons of the central nervous system regulate the synthesis of the main developmental hormones. Neuropeptides involved in this neuroendocrine cascade have been identified in lepidopterans and dictyopterans. Since these organisms are not suitable for genetic research, we identified peptidergic brain neurons innervating the ring gland in Drosophila melanogaster. In larvae of Drosophila, ecdysteroids and juvenile hormones are produced by the ring gland, which is composed of the prothoracic gland, the corpus allatum, and the corpora cardiaca. Using the GAL4 enhancer trap system, we mapped those neurons of the central nervous system that innervate the ring gland. Eleven groups of neurosecretory neurons and their target tissues were identified. Five neurons of the lateral protocerebrum directly innervate the prothoracic gland or corpus allatum cells of the ring gland and are believed to regulate ecdysteroid and juvenile hormone titers. Axons of the circadian pacemaker neurons project onto dendritic fields of these five neurons. This connection might be the neuronal substrate of the circadian rhythms of molting and metamorphosis in Drosophila. Most of the neurons presented here have not been described before. The enhancer trap lines labeling them will be valuable tools for the analysis of neuronal as well as genetic regulation in insect development.

Animals↗
PubMed2000

Domina (Dom), a new Drosophila member of the FKH/WH gene family, affects morphogenesis and is a suppressor of position-effect variegation.

Domina (Dom) is a novel member of the FKH/WH transcription factor gene family of Drosophila. Two alternatively polyadenylated Dom transcripts of 2.9 and 3.9 kb encode a 719-amino-acid protein with a FKH/WH domain and a putative acidic transactivation domain. Dom is mainly expressed in the central and peripheral nervous system. Homozygous mutants show rough eyes, irregular arrangement of bristles, extended wings, defective posterior wing margins, and a severely diminished vitality and fertility. Heterozygous Dom flies are morphologically wild type but show suppression of position-effect variegation. Consistently with this chromatin effect DOM protein is accumulated in the chromocenter and, as expected from a transcription factor, is found at specific euchromatic loci. Sequence comparison suggests that DOM of Drosophila is homologous to the chordate WHN proteins. The chromatin modifying capability of DOM is probably based on the FKH/WH domain, which shows a remarkable structural similarity to the winged-helix structures of H1 and the central globular domain of H5.

Alleles↗
PubMed1999

The helix-loop-helix proteins dAP-4 and daughterless bind both in vitro and in vivo to SEBP3 sites required for transcriptional activation of the Drosophila gene Sgs-4.

The expression of Sgs genes in the salivary gland of the third instar larva of Drosophila is a spatially restricted response to signalling by the steroid hormone 20-hydroxyecdysone. For Sgs-4, we have previously demonstrated that its strictly tissue and stage-specific expression is the result of combined action of the ecdysone receptor and secretion enhancer binding proteins (SEBPs). One of these SEBPs, SEBP2, was shown to be the product of the homeotic gene fork head. Together with SEBP3, SEBP2 appears to be responsible for the spatial restriction of the hormone response of Sgs-4. Here, we show that SEBP3 is a heterogeneous binding activity that consists of different helix-loop-helix (HLH) proteins. We cloned the Drosophila homologue of human transcription factor AP-4 (dAP-4) and identified it as one of these HLH proteins. The dAP-4 protein shows great similarity to its human and Caenorhabditis counterparts within the bHLHZip domain, the second leucine zipper dimerization motif, and a third region of unknown function. The expression pattern of dAP-4 indicates that it is a ubiquitously expressed HLH protein in Drosophila. As a second component of SEBP3 we identified the Daughterless (Da) protein, which is also ubiquitously expressed and binds to SEBP3 sites independent of dAP-4. Since both dAP-4 and Da can be detected in situ at transposed Sgs-4 transcriptional control elements in polytene salivary gland chromosomes, we propose that each of the two proteins contributes to the transcriptional control of Sgs-4.

Amino Acid Sequence↗
PubMed1999

Structure and regulation of the salivary gland secretion protein gene Sgs-1 of Drosophila melanogaster.

The Drosophila melanogaster gene Sgs-1 belongs to the secretion protein genes, which are coordinately expressed in salivary glands of third instar larvae. Earlier analysis had implied that Sgs-1 is located at the 25B2-3 puff. We cloned Sgs-1 from a YAC covering 25B2-3. Despite using a variety of vectors and Escherichia coli strains, subcloning from the YAC led to deletions within the Sgs-1 coding region. Analysis of clonable and unclonable sequences revealed that Sgs-1 mainly consists of 48-bp tandem repeats encoding a threonine-rich protein. The Sgs-1 inserts from single lambda clones are heterogeneous in length, indicating that repeats are eliminated. By analyzing the expression of Sgs-1/lacZ fusions in transgenic flies, cis-regulatory elements of Sgs-1 were mapped to lie within 1 kb upstream of the transcriptional start site. Band shift assays revealed binding sites for the transcription factor fork head (FKH) and the factor secretion enhancer binding protein 3 (SEBP3) at positions that are functionally relevant. FKH and SEBP3 have been shown previously to be involved in the regulation of Sgs-3 and Sgs-4. Comparison of the levels of steady state RNA and of the transcription rates for Sgs-1 and Sgs-1/lacZ reporter genes indicates that Sgs-1 RNA is 100-fold more stable than Sgs-1/lacZ RNA. This has implications for the model of how Sgs transcripts accumulate in late third instar larvae.

Amino Acid Sequence↗
PubMed1998

The pipsqueak protein of Drosophila melanogaster binds to GAGA sequences through a novel DNA-binding domain.

Pipsqueak (Psq) belongs to a family of proteins defined by a phylogenetically old protein-protein interaction motif. Like the GAGA factor and other members of this family, Psq is an important developmental regulator in Drosophila, having pleiotropic functions during oogenesis, embryonic pattern formation, and adult development. The GAGA factor controls the transcriptional activation of homeotic genes and other genes by binding to control elements containing the GAGAG consensus motif. Binding is associated with formation of an open chromatin structure that makes the control regions accessible to transcriptional activators. We show here that Psq contains a novel DNA-binding domain, which binds, like the GAGA factor zinc finger DNA-binding domain, to target sites containing the GAGAG consensus motif. Binding is suppressed, as in the GAGA factor and other proteins of the family, by the associated protein-protein interaction motif. The DNA-binding domain, which we call the Psq domain, is identical with a previously identified region consisting of four tandem repeats of a conserved 50-amino acid sequence, the Psq motif. The Psq domain seems to be structurally related to known DNA-binding domains, both in its repetitive character and in the putative three-alpha-helix structure of the Psq motif, but it lacks the conserved sequence signatures of the classical eukaryotic DNA-binding motifs. Psq may thus represent the prototype of a new family of DNA-binding proteins.

Amino Acid Sequence↗
PubMed1998

The mub gene encodes a protein containing three KH domains and is expressed in the mushroom bodies of Drosophila melanogaster.

The ring gland function of Drosophila melanogaster is controlled by the CNS. To identify genes that are active in brain cells and are involved in the ring gland control, we analysed enhancer trap lines with respect to CNS- and/or ring gland-specific lacZ expression in third-instar larvae. From one of the enhancer trap lines, which shows specific lacZ expression in the CNS and prothoracic part of the ring gland, the mub gene was cloned. The gene is strongly expressed in the mushroom bodies throughout development. Nucleotide sequence analysis of cDNA clones revealed a high degree of similarity to vertebrate RNA binding KH domain proteins, suggesting a function of the MUB protein in binding and stabilizing of specific mRNAs in the mushroom bodies. Null mutants of the mub gene do not exhibit a visible mutant phenotype. We speculate, therefore, that the mub gene is involved in learning and memory processes.

Amino Acid Sequence↗
PubMed1998

Comparison of the GAGA factor genes of Drosophila melanogaster and Drosophila virilis reveals high conservation of GAGA factor structure beyond the BTB/POZ and DNA-binding domains.

As a member of the trithorax-group, the Trithorax-like (Trl) gene of Drosophila melanogaster contributes to the expression of homeotic genes and many other genes. Trl encodes different isoforms of the GAGA factor which is thought to act as an "antirepressor" of transcription by remodelling chromatin structure and thereby rendering control regions accessible for transcriptional activators. A more global role of the GAGA factor in chromatin structure and function is suggested by various phenotypes of Trl mutations, such as modification of position effect variegation. To better define the molecular basis of these pleiotropic effects, we cloned cDNAs encoding the GAGA isoforms of D. melanogaster and a distantly related species, D. virilis. We also characterized the genomic organization of both the D. melanogaster and D. virilis genes, and analysed the expression patterns of isoform-specific mRNAs. The D. virilis GAGA isoforms show high similarity to their D. melanogaster counterparts, particularly within the BTB/POZ protein-interaction and the zinc finger DNA-binding domains. Interestingly, conservation clearly extends beyond the previously defined limits of these domains. Moreover, the comparison reveals a completely conserved block of amino acid residues located between the BTB/POZ and DNA-binding domains, and a high conservation of the C-terminus specific for one of the GAGA isoforms. Thus, sequences of as yet unknown functions are defined as rewarding targets for further mutational analyses. The high conservation of the GAGA proteins of the two species is in accord with the nearly identical genomic organization and expression patterns of the corresponding genes.

Amino Acid Sequence↗
PubMed1997

Two new regulatory elements controlling the Drosophila Sgs-3 gene are potential ecdysone receptor and fork head binding sites.

We identified two regulatory elements in the upstream region of the Drosophila Sgs-3 gene which are both able to bind the ecdysone receptor (EcR/USP) and the product of the fork head gene. Interestingly, only one of the EcR/USP binding sites is able to recognize in vitro-translated EcR/USP, which provides evidence for the existence of different receptor forms having different DNA binding specificities. Deletions of the elements lead to a reduced accumulation of Sgs-3 mRNA without altering the temporal expression profile of the gene. The data are consistent with the hypothesis that the ecdysone receptor directly contributes to the transcriptional activation of Sgs-3 by binding to at least one of the two elements. Since also the Sgs-4 gene is controlled by a functional EcR/USP binding site, a direct participation of EcR/USP in the formation of regulatory complexes may be of general importance for the hormonal control of Sgs genes.

Animals↗
PubMed1997

Clonning and sequence analysis of the 26 kDa glutathiones-transferase gene of Schistosoma mekongi.

The number of genomic DNA or cDNA sequences of Schistosoma mekongi accessible in Genbank or EMBL is very limited up to now. Recently, two reports have appeared on the molecular phylogeny of Schistosoma species inferred from partial sequence data of rRNA genes; no further sequence data of S. mekongi is available yet. Knowledge of the molecular structure of protein coding genes of S. mekongi will provide a better understanding of gene function in the genus Schistosoma. A cDNA library of S. mekongi adult male was constructed and a cDNA encoding the 26 kDa glutathione S-transferase protein of this species was cloned. Sequence analysis of this cDNA confirmed the close phylogenetic relationship of S. mekongi to S. japonicum.

Amino Acid Sequence↗
PubMed1996

The fork head product directly specifies the tissue-specific hormone responsiveness of the Drosophila Sgs-4 gene.

Here we describe the identification of four binding sites of secretion enhancer binding protein 2 (SEBP2) in the regulatory region of the Drosophila salivary gland secretion protein gene 4 (Sgs-4) and show that despite these sites' correspondence with previously described Broad-Complex protein binding sites, SEBP2 is a Broad-Complex-independent factor encoded by the region-specific homeotic gene fork head (fkh). Two of the Fork head/SEBP2 binding sites are located within an ecdysone response unit which controls the tissue- and stage-specific responses of Sgs-4 to the steroid hormone 20-hydroxyecdysone. We demonstrate that these binding sites are relevant to the transcriptional activation of Sgs-4 and show that Fork head also binds to the Sgs-4 ecdysone response unit in vivo. Aside from being involved in the control of decisions during embryonic development, fkh thus participates directly in the control of specialized functions of differentiated cells at later stages of development.

Animals↗
PubMed1995

Ecdysone regulation of the Drosophila Sgs-4 gene is mediated by the synergistic action of ecdysone receptor and SEBP 3.

The steroid hormone 20-hydroxyecdysone controls both induction and repression of the Drosophila 'intermolt gene' Sgs-4. We show here that the ecdysone receptor binds to two sites, element I and element II, in the regulatory region of Sgs-4. A functional analysis revealed that element II appears to be of no importance for Sgs-4 expression, while element I proved to be an ecdysone response element that is necessary, but not sufficient, for induction of Sgs-4 expression. Our results provide no evidence that repression of Sgs-4 expression is mediated by one of the two receptor binding sites. In the close vicinity of elements I and II, we detected two binding sites of secretion enhancer binding protein 3 (SEBP 3). Like receptor element I, one of these sites also proved to be necessary, but not sufficient, for expression of Sgs-4. Therefore, induction of Sgs-4 requires binding of both ecdysone receptor and SEBP 3 to a complex hormone response unit, which also contains binding sites for a third factor, SEBP 2. The SEBP 2 sites coincide with binding sites of products of the Broad-Complex locus, which has been implicated recently with transduction of the hormonal signal. Thus, the available data suggest that induction of Sgs-4, and possibly other 'intermolt genes', is a combination of a primary and a secondary response to the hormone.

Animals↗
PubMed1994

The protein encoded by the Drosophila position-effect variegation suppressor gene Su(var)3-9 combines domains of antagonistic regulators of homeotic gene complexes.

Modifier mutations of position-effect variegation (PEV) represent a useful tool for a genetic and molecular dissection of genes connected with chromatin regulation in Drosophila. The Su(var)3-9 gene belongs to the group of haplo suppressor loci which manifest a triplo enhancer effect. Mutations show a strong suppressor effect even in the presence of PEV enhancer mutations, indicating a central role of this gene in the regulation of PEV. By molecular analysis, Su(var)3-9 could be correlated with a 2.4 kb transcript which encodes a putative protein of 635 amino acids containing a chromo domain and a region of homology to Enhancer of zeste and trithorax, two antagonistic regulators of the Antennapedia and Bithorax gene complexes, as well as to the human protein ALL-1/Hrx which is implicated in acute leukemias. This region of homology is found in all four proteins at the C-terminus. The homology of Su(var)3-9 to both negative (Polycomb and Enhancer of zeste) and positive (trithorax) regulators of the Antennapedia and Bithorax complexes also suggests similarities in the molecular processes connected with stable transmission of a determined state and the clonal propagation of heterochromatinization.

Amino Acid Sequence↗
PubMed1993

P transposon-induced dominant enhancer mutations of position-effect variegation in Drosophila melanogaster.

P transposon induced modifier mutations of position-effect variegation (PEV) were isolated with the help of hybrid dysgenic crosses (pi 2 strain) and after transposition of the mutator elements pUChsneory+ and P[lArB]. Enhancer mutations were found with a ten times higher frequency than suppressors. The 19 pUChsneory(+)- and 15 P[lArB]-induced enhancer mutations can be used for cloning of genomic sequences at the insertion sites of the mutator elements via plasmid rescue. Together with a large sample of X-ray-induced (48) and spontaneous (93) enhancer mutations a basic genetic analysis of this group of modifier genes was performed. On the basis of complementation and mapping data we estimate the number of enhancer genes at about 30 in the third chromosome and between 50 and 60 for the whole autosome complement. Therefore, enhancer of PEV loci are found in the Drosophila genome as frequently as suppressor genes. Many of the enhancer mutations display paternal effects consistent with the hypothesis that some of these mutations can induce genomic imprinting. First studies on the developmentally regulated gene expression of PEV enhancer genes were performed by beta-galactosidase staining in P[lArB] induced mutations.

Animals↗
PubMed1990

Two puff-specific proteins bind within the 2.5 kb upstream region of the Drosophila melanogaster Sgs-4 gene.

The Drosophila nuclear proteins Bj6 and Bx42 characterized previously are detected in a series of developmentally active puffs on salivary gland chromosomes. Here the binding of both proteins at puff 3C11-12 containing the glue protein gene Sgs-4 is described in more detail. By deletion analysis we show that both proteins bind within a chromosomal segment containing 17-19 kb of DNA surrounding the Sgs-4 gene. They are detectable at this site during the intermoult stages, before the puff regresses in response to the moulting hormone ecdysone. If the Sgs-4 gene together with flanking DNA sequences is brought into a different chromosomal position by P element transfer, both proteins are detected at this new location. Both proteins are bound to the chromosome within the range of 2.5 kb DNA upstream of the Sgs-4 gene. A strain containing a 52 bp deletion within this region fails to bind Bx42 protein suggesting that the missing DNA, which overlaps a hypersensitive region, may be required for the binding of the Bx42 protein.

Animals↗
PubMed1990

Promoter is an important determinant of developmentally regulated puffing at the Sgs-4 locus of Drosophila melanogaster.

Sgs-4 is one of the eight known genes coding for larval secretion proteins in Drosophila melanogaster. High-level transcription of the endogenous Sgs genes in salivary glands is accompanied by chromosome puffing at the Sgs gene loci. Naturally occurring mutations of the Sgs-4 promoter region diminish both the level of Sgs-4 expression and the puff size; in null-producers no puff is formed. P element-mediated transformation experiments were performed to clarify this apparent causal relation between transcription and puffing. Sgs-4 upstream sequences, unchanged or recombined with sequences from differently expressed alleles, were fused with Sgs-4 coding and downstream sequences or with the coding sequence of the viral oncogene v-mil. Analyses of the expression of these fragments at the RNA and protein levels and of their capacity for puff formation demonstrate uncoupling of transcription and puffing. That is, high-level transcription is independent of chromosome puffing and does not necessarily induce puffing, and developmentally regulated chromosome puffing is independent of significant transcriptional activity within the puff. Our results show that the strength of the Sgs-4 promoter located within the upstream region from -1 to -840 determines the formation of a puff. No specific effects could be detected on either transcription or puffing by decondensed versus compact chromatin adjoining the transposed DNA at the sites of insertion in transformants. A model in which trans-acting factors binding to the promoter region initiate puffing is proposed.

Animals↗
PubMed1987

Upstream sequences of dosage-compensated and non-compensated alleles of the larval secretion protein gene Sgs-4 in Drosophila.

The X chromosomal gene Sgs-4 coding for a larval secretion protein of Drosophila melanogaster is expressed stage and tissue specifically and is hyperexpressed in male larvae of most Drosophila stocks which show dosage compensation. We analysed three Sgs-4 alleles which differ in the size of their coding region, in the intensity of their expression and in the level of dosage compensation. The size and amount of Sgs-4 proteins directly reflect those of RNAs. Different RNA sizes result from different numbers of 21 bp repeats within the structural genes. We sequenced 2.8 kb of DNA upstream of the transcription initiation site of the three alleles. Sequences known to be essential for correct gene expression were located. The only difference within DNA sequences from -1 to -1200 between two alleles with different degrees of expression and differing in dosage compensation is a C to T transition at -344 within a supposed consensus sequence for ecdysone receptor complex binding (ECR). This mutation is partly located within a region of dyad symmetry. Alleles with identical expression show identical mutations within a GTT-rich region at -1.2 kb, but differ within a GT-rich region at -2.0 kb. A polyadenylated 0.5 kb RNA was found to be transcribed from the GTT-rich region of the strand opposite to that of Sgs-4. The corresponding gene is active only in larval salivary glands and, therefore, is named gland specific gene, gsg.

Alleles↗