-
Developmental Biology Aug 2006A key step in organogenesis of the Drosophila tracheal system is the integration of isolated tracheal metameres into a connected tubular network. The interaction of...
A key step in organogenesis of the Drosophila tracheal system is the integration of isolated tracheal metameres into a connected tubular network. The interaction of tracheal cells with surrounding mesodermal cells is crucial in this process. In particular, single mesodermal cells called bridge-cells are essential for the guided outgrowth of dorsal trunk branches to direct formation of the main airway, the dorsal trunk. Here, we present evidence that the two leucine-rich repeat transmembrane proteins Capricious and Tartan contribute differently to the formation of branch interconnections during tracheal development. Capricious is specifically localized on the surface of bridge-cells and facilitates the outgrowing dorsal trunk cells of adjacent metameres toward each other. We show that Capricious requires both extracellular and intracellular domains during tracheal branch outgrowth. In contrast, Tartan is expressed broadly in mesodermal cells and exerts its role in tracheal branch outgrowth through its extracellular domain. We propose that Capricious contributes to the instructive role of bridge-cells whereas Tartan provides permissive substrate for the migrating tracheal cells during the network formation.
Topics: Amino Acid Sequence; Animals; Cell Movement; Drosophila Proteins; Drosophila melanogaster; Leucine; Leucine-Rich Repeat Proteins; Membrane Proteins; Molecular Sequence Data; Proteins; Trachea
PubMed: 16764850
DOI: 10.1016/j.ydbio.2006.04.462 -
Genetics Nov 2019Insect odorant-binding proteins (OBPs) are a large, diverse group of low-molecular weight proteins secreted into the fluid bathing olfactory and gustatory neuron...
Insect odorant-binding proteins (OBPs) are a large, diverse group of low-molecular weight proteins secreted into the fluid bathing olfactory and gustatory neuron dendrites. The best-characterized OBP, LUSH (OBP76a) enhances pheromone sensitivity enabling detection of physiological levels of the male-specific pheromone, 11- vaccenyl acetate. The role of the other OBPs encoded in the genome is largely unknown. Here, using clustered regularly interspaced short palindromic repeats/Cas9, we generated and characterized the loss-of-function phenotype for two genes encoding homologous OBPs, OS-E (OBP83b) and OS-F (OBP83a). Instead of activation defects, these extracellular proteins are required for normal deactivation of odorant responses to a subset of odorants. Remarkably, odorants detected by the same odorant receptor are differentially affected by the loss of the OBPs, revealing an odorant-specific role in deactivation kinetics. In stark contrast to mutants, the mutants have normal activation kinetics to the affected odorants, even at low stimulus concentrations, suggesting that these OBPs are not competing for these ligands with the odorant receptors. We also show that and are functionally redundant as either is sufficient to revert the mutant phenotype in transgenic rescue experiments. These findings expand our understanding of the roles of OBPs to include the deactivation of odorant responses.
Topics: Animals; Carrier Proteins; Drosophila Proteins; Drosophila melanogaster; Loss of Function Mutation; Phenotype; Protein Binding
PubMed: 31492805
DOI: 10.1534/genetics.119.302629 -
Methods in Molecular Biology (Clifton,... 2022Asymmetry in the migrating group of cells is critical for efficient directed movement observed in normal development and in pathological conditions like tumor cell...
Asymmetry in the migrating group of cells is critical for efficient directed movement observed in normal development and in pathological conditions like tumor cell metastasis. This is conspicuously detected at the level of polarized protrusions and differential localization of various polarity proteins in collectively moving clusters. Over the years, border cell migration in Drosophila oogenesis has emerged as an excellent model system for studying polarity in the migrating group of cells. Here we report two protocols employing live cell imaging and tissue immunohistochemistry to evaluate the polarity in migrating border cell clusters.
Topics: Animals; Cell Movement; Cell Polarity; Drosophila; Drosophila Proteins; Drosophila melanogaster; Oogenesis
PubMed: 35147959
DOI: 10.1007/978-1-0716-2035-9_28 -
The Journal of Cell Biology Mar 2018Growth of epithelial tissues is regulated by a plethora of components, including signaling and scaffolding proteins, but also by junctional tension, mediated by the...
Growth of epithelial tissues is regulated by a plethora of components, including signaling and scaffolding proteins, but also by junctional tension, mediated by the actomyosin cytoskeleton. However, how these players are spatially organized and functionally coordinated is not well understood. Here, we identify the scaffolding protein Big bang as a novel regulator of growth in epithelial cells of the wing disc by ensuring proper junctional tension. Loss of results in the reduction of the regulatory light chain of nonmuscle myosin, Spaghetti squash. This is associated with an increased apical cell surface, decreased junctional tension, and smaller wings. Strikingly, these phenotypic traits of mutant discs can be rescued by expressing constitutively active Spaghetti squash. Big bang colocalizes with Spaghetti squash in the apical cytocortex and is found in the same protein complex. These results suggest that in epithelial cells of developing wings, the scaffolding protein Big bang controls apical cytocortex organization, which is important for regulating cell shape and tissue growth.
Topics: Animals; Drosophila Proteins; Drosophila melanogaster; Epithelial Cells; Gene Expression Regulation, Developmental; Imaginal Discs; Wings, Animal
PubMed: 29326288
DOI: 10.1083/jcb.201705104 -
Cell Aug 2007GRASP proteins associate with the Golgi apparatus and have been implicated in the stacking of Golgi cisternae, vesicle tethering, and mitotic progression, but their...
GRASP proteins associate with the Golgi apparatus and have been implicated in the stacking of Golgi cisternae, vesicle tethering, and mitotic progression, but their specific functions are unclear. In this issue, Kinseth et al. (2007) show unexpectedly that a GRASP homolog is required for an unconventional secretory pathway that bypasses the usual route for Golgi-dependent membrane traffic.
Topics: Animals; Dictyostelium; Drosophila Proteins; Golgi Apparatus; Golgi Matrix Proteins; Membrane Proteins; Signal Transduction
PubMed: 17693251
DOI: 10.1016/j.cell.2007.07.030 -
Nucleic Acids Research Feb 2021In arthropods, zinc finger-associated domains (ZADs) are found at the N-termini of many DNA-binding proteins with tandem arrays of Cys2-His2 zinc fingers (ZAD-C2H2...
In arthropods, zinc finger-associated domains (ZADs) are found at the N-termini of many DNA-binding proteins with tandem arrays of Cys2-His2 zinc fingers (ZAD-C2H2 proteins). ZAD-C2H2 proteins undergo fast evolutionary lineage-specific expansion and functional diversification. Here, we show that all ZADs from Drosophila melanogaster form homodimers, but only certain ZADs with high homology can also heterodimerize. CG2712, for example, is unable to heterodimerize with its paralog, the previously characterized insulator protein Zw5, with which it shares 46% homology. We obtained a crystal structure of CG2712 protein's ZAD domain that, in spite of a low sequence homology, has similar spatial organization with the only known ZAD structure (from Grauzone protein). Steric clashes prevented the formation of heterodimers between Grauzone and CG2712 ZADs. Using detailed structural analysis, site-directed mutagenesis, and molecular dynamics simulations, we demonstrated that rapid evolutionary acquisition of interaction specificity was mediated by the more energy-favorable formation of homodimers in comparison to heterodimers, and that this specificity was achieved by multiple amino acid substitutions resulting in the formation or breaking of stabilizing interactions. We speculate that specific homodimerization of ZAD-C2H2 proteins is important for their architectural role in genome organization.
Topics: Animals; Crystallography, X-Ray; DNA-Binding Proteins; Dimerization; Drosophila; Drosophila Proteins; Drosophila melanogaster; Models, Molecular; Mutagenesis; Protein Multimerization; Transcription Factors; Zinc Fingers
PubMed: 33638995
DOI: 10.1093/nar/gkab061 -
Acta Crystallographica. Section F,... Apr 2014Drosophila melanogaster contains two calcium-binding proteins, Frq1 and Frq2, in the nervous system that control the number of synapses and the probability of release....
Drosophila melanogaster contains two calcium-binding proteins, Frq1 and Frq2, in the nervous system that control the number of synapses and the probability of release. To understand the differential function of the two proteins, whose sequence is only 5% dissimilar, the crystal structures of Frq1 and Frq2 are needed. Here, the cloning, expression, purification, crystallization and preliminary crystallographic analysis of Frq2 are presented. The full-length protein was purified using a two-step chromatographic procedure. Two different diffracting crystal forms were obtained using a progressive streak-seeding method and detergents.
Topics: Animals; Calcium-Binding Proteins; Cloning, Molecular; Crystallization; Crystallography, X-Ray; Drosophila Proteins; Drosophila melanogaster
PubMed: 24699756
DOI: 10.1107/S2053230X14005408 -
EMBO Reports Apr 2016might be a suitable tool for drug discovery, but it is not clear how often the compounds identified will work in humans. A detailed look at previous work suggests that...
might be a suitable tool for drug discovery, but it is not clear how often the compounds identified will work in humans. A detailed look at previous work suggests that compounds active in both species might be relatively common. [Image: see text]
Topics: Animals; Drosophila; Drosophila Proteins; Drug Discovery; Humans; Models, Animal; Proteomics; Translational Research, Biomedical
PubMed: 26882560
DOI: 10.15252/embr.201642080 -
Current Opinion in Structural Biology Oct 2007Notch receptors are approximately 300 kDa cell surface glycoproteins whose activation by Notch ligands regulates cell fate decisions in the metazoa. The extracellular... (Review)
Review
Notch receptors are approximately 300 kDa cell surface glycoproteins whose activation by Notch ligands regulates cell fate decisions in the metazoa. The extracellular domain of Notch receptors has many epidermal growth factor like repeats that are glycosylated with O-fucose and O-glucose glycans as well as N-glycans. Disruption of O-fucose glycan synthesis leads to severe Notch signaling defects in Drosophila and mammals. Removal or addition of O-fucose glycan consensus sites on Notch receptors also leads to Notch signaling defects. Ligand binding and ligand-induced Notch signaling assays have provided insights into how changes in the O-fucose glycans of Notch receptors alter Notch signaling.
Topics: Animals; Carbohydrate Sequence; Drosophila; Drosophila Proteins; Fucose; Genes, Insect; Glycosylation; Humans; Mammals; Models, Biological; Molecular Sequence Data; Mutation; N-Acetylglucosaminyltransferases; Receptors, Notch; Signal Transduction
PubMed: 17964136
DOI: 10.1016/j.sbi.2007.09.007 -
Genome Biology 2007A recent report describes the identification through the use of in vitro selection of a peptide that antagonizes Methuselah signaling in Drosophila in vitro and extends... (Review)
Review
A recent report describes the identification through the use of in vitro selection of a peptide that antagonizes Methuselah signaling in Drosophila in vitro and extends fly life span in vivo.
Topics: Animals; Drosophila Proteins; Drosophila melanogaster; Drug Design; Ligands; Longevity; Oligopeptides; Protein Conformation; Receptors, G-Protein-Coupled
PubMed: 17764591
DOI: 10.1186/gb-2007-8-8-222