ABSTRACT
Leg-arista-wing complex (lawc) mutations affect the expression of D. melanogaster proteins homologous to a human basic transcription factor, TBP (TATA-box binding protein) Related Factor 2 (TRF2), specifically involved in development. The paper for the first time reports the application of genetic screens for various genomic regions to recover genetic interactions between the lawc/Trf2 gene and other genes and genetic loci by using Deficiency Kit lines with small deletions in total providing maximal coverage of the genome. The deletion mapping allowed us to recover 26 genomic regions that, when deleted, are lethal or modify the mutant phenotype due to a decreased TRF2 expression level. These deletions could be useful in identifying both novel TRF2 targets and its positive and negative regulators. There is evidence that TRF2 can be a component of high molecular DNA Replication-related Element Factor (DREF)- and Nucleosome Remodeling Factor (NURF)-containing complexes. The present study for the first time reports new genetic interactions of lawc/Trf2 with genes that encode basic and specific transcription factors. In most cases, if mutated, those genes caused developmental defects or death of progeny. However, in the case of the e(y) 1 gene, coding for the Taf9 transcription factor, only the male reproductive system is impaired when the lawc/Trf2 phenotype is associated with a e(y)l gene mutation. Mutant lawc(p1)e(y)1(u1) males become infertile due to primary spermatocyte maturation arrest and impaired premeiotic chromosome condensation in germ cells.
Subject(s)
Drosophila Proteins/genetics , Drosophila melanogaster/growth & development , Drosophila melanogaster/genetics , Gene Expression Regulation, Developmental , Telomeric Repeat Binding Protein 2/genetics , Amino Acid Sequence , Animals , Chromosomes, Insect , Drosophila Proteins/metabolism , Genomics/methods , Male , Molecular Sequence Data , Mutation , Spermatocytes/metabolism , TATA-Binding Protein Associated Factors/genetics , TATA-Binding Protein Associated Factors/metabolism , Telomeric Repeat Binding Protein 2/metabolism , Transcription Factor TFIID/genetics , Transcription Factor TFIID/metabolism , Transcription Factors/genetics , Transcription Factors/metabolismABSTRACT
The influence of the P-element built into the area of the CG5017 gene on the mutation of the spineless (ss) gene was studied. It was shown that the insertion of the P-element decreased the level of transcription of CG5017 approximately twofold. Modulation of the level of transcription of the CG5017 gene helped demonstrate, for the first time, its influence on the phenotypic manifestation of the mutation of the ss gene, which shows their interaction in the process of regulation of morphogenesis of limbs in Drosophila melanogaster.
Subject(s)
Drosophila Proteins/metabolism , Embryo, Nonmammalian/embryology , Extremities/embryology , Morphogenesis/physiology , Receptors, Aryl Hydrocarbon/metabolism , Animals , Drosophila Proteins/genetics , Drosophila melanogaster , Receptors, Aryl Hydrocarbon/geneticsABSTRACT
Preliminary analysis of the leg-arista-wing complex (lawc) gene region in the corresponding mutants revealed P element insertion in the transcribed region of this locus. To demonstrate the main role of P element insertion in the complex pleiotropic phenotypic manifestation of the lawc gene, a system using P-Ph chimeric protein and based on the P-mediated repression of the lawc in vivo transcription was applied. As a result, extreme lawc-mutant phenotypes were obtained and examined. The P-Ph-mediated decrease of the level of the lawc gene transcription was also demonstrated.
Subject(s)
Drosophila/genetics , Gene Expression Regulation/physiology , Genes, Insect , Genes, Suppressor , Recombinant Fusion Proteins/metabolism , Alleles , Animals , Drosophila/metabolism , Insect Proteins/genetics , Insect Proteins/metabolism , Mutagenesis, Insertional , Mutation , Phenotype , Transcription, GeneticSubject(s)
HSP70 Heat-Shock Proteins/pharmacology , Neuroglia/pathology , Neurons/transplantation , Transplantation, Heterologous/pathology , Animals , Cicatrix/pathology , Drosophila melanogaster/embryology , Embryo, Nonmammalian/cytology , Embryo, Nonmammalian/physiology , Graft Survival/physiologyABSTRACT
The influence of xenografts of Drosophila melanogaster embryonic nerve cells on the development of embryonic neurohomografts in the adult rat brain has been investigated. Embryonic nerve cells, marked with bacterial galactosidase gene (lacZ) from D. melanogaster strain with a mutation in the Delta locus, were transplanted into adult rat brain. Drosophila cells were easily identifiable in brain histological sections by X-gal staining. Xenografts survived for at least 2-3 weeks in the recipient brain after the operation to be subsequently attacked by macrophages. Importantly, no glial scar was formed around the xenograft. The addition of Drosophila embryonic nerve cells to a homograft of rat embryonic neural tissue facilitated the survival and development of this homograft by blocking the glial scar formation, stimulating vascularization of the graft area and differentiation of the implanted embryonic nerve cells.
Subject(s)
Brain/surgery , Drosophila melanogaster/embryology , Fetal Tissue Transplantation , Nerve Tissue/transplantation , Neuroglia/cytology , Transplantation, Heterologous , Animals , Drosophila melanogaster/genetics , Embryo, Nonmammalian/cytology , Gene Transfer Techniques , Graft Survival , Lac Operon/genetics , Mutation , Nerve Tissue Proteins/genetics , Rats , beta-Galactosidase/geneticsABSTRACT
Transgenic Drosophila stocks were selected. These stocks contained human ngf and bdnf genes under Drosophila heat-shock promoter. The expression of foreign genes during Drosophila development was demonstrated with Northern and whole mount hybridization.
Subject(s)
Brain-Derived Neurotrophic Factor/genetics , Drosophila melanogaster/embryology , Gene Expression Regulation, Developmental , Nerve Growth Factors/genetics , Animals , Animals, Genetically Modified , Brain-Derived Neurotrophic Factor/biosynthesis , Drosophila melanogaster/genetics , Drosophila melanogaster/metabolism , Embryo, Nonmammalian , Heat-Shock Proteins/genetics , Humans , Molecular Probe Techniques , Nerve Growth Factors/biosynthesis , Promoter Regions, Genetic , TransgenesSubject(s)
Animals, Genetically Modified/genetics , Drosophila melanogaster/genetics , Gene Expression/genetics , Nerve Growth Factors , Nerve Tissue Proteins/genetics , Animals , Blotting, Northern/methods , Blotting, Southern/methods , Embryo, Nonmammalian/chemistry , Glial Cell Line-Derived Neurotrophic Factor , Humans , In Situ Hybridization/methods , Larva/chemistry , Larva/genetics , Microinjections/methods , Neurons/metabolismABSTRACT
A transgenic line of Drosophila melanogaster was selected which carried the following genes: Delta, lacZ (for bacterial galactosidase), and human GDNF (for glial cell line-derived neurotrophic factor). Drosophila neuroectodermal embryonic cells were transplanted with the embryonic neurohomografts into the occipital brain region of an adult rat. Xenografts were found to block scar formation at the graft-host tissue boundary, stimulated homograft development (so that it was twice as large as the control homograft transplanted alone with no xenograft added), and noticeably improved vascularization of the homograft area.
Subject(s)
Brain/anatomy & histology , Drosophila melanogaster/genetics , Neocortex/transplantation , Nerve Growth Factors , Nerve Tissue Proteins/genetics , Animals , Brain/surgery , Cell Transplantation , Drosophila melanogaster/cytology , Drosophila melanogaster/embryology , Embryo, Nonmammalian/cytology , Female , Fetal Tissue Transplantation , Gene Transfer Techniques , Glial Cell Line-Derived Neurotrophic Factor , Humans , Immunohistochemistry , Intracellular Signaling Peptides and Proteins , Lac Operon , Membrane Proteins/genetics , Neocortex/embryology , Rats , Selection, Genetic , Transplantation, HeterologousABSTRACT
Due to the complete absence of ribosomal DNA (genetic symbol bb-), the Xbb- chromosome of Drosophila is lethal both in homozygous conditions and in compound with the Xbb- chromosome. However, in the cross between the C(1)RM/Ybb- females and the Xbb-/BSYbb+ males, characterized by the development of lethal Xbb-/Ybb- zygotes, two fertile males were detected. These males possessed all the markers of the Xbb- chromosome but lacked the Y chromosome BS marker. Genetic analysis of their progeny showed that genes responsible for restoration of viability and fertility of these exceptional males were associated with the X chromosome. The crossover tests showed that in one case these genes were tightly linked to the w locus (the bbAM1 allele), and in the second case they were located 12.6 map units to the right of the Tu locus (the bbAM7 allele). It has also been shown that the bb locus was transposed to the X chromosome within the short arm of Y chromosome. Transposition of the BSYbb+ chromosome-specific rDNA sequences to the X chromosome was confirmed by means of Southern blotting. These data indicate that replacement of the bb locus is realized by transposition rather than recombination.