-
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 -
Tissue Barriers Apr 2017The role of Slit/Robo signaling has extended from initial axon repulsion in the developing nervous system to organ morphogenesis, cancer development and angiogenesis.... (Review)
Review
The role of Slit/Robo signaling has extended from initial axon repulsion in the developing nervous system to organ morphogenesis, cancer development and angiogenesis. Slit/Robo signaling regulates similar pathways within these processes. Slit/Robo ensures the homeostasis of the dynamic interaction between cell-cell and cell-matrix interactions. The dysregulation of Slit/Robo signaling damages the tissue barrier, resulting in developmental abnormalities or disease. Here, we summarize how Slit/Robo controls kidney morphogenesis and describe the dual roles of Slit/Robo signaling in the regulation of tumorigenesis and angiogenesis.
Topics: Animals; Cell Communication; Drosophila Proteins; Models, Biological; Nerve Tissue Proteins; Receptors, Immunologic; Signal Transduction; Roundabout Proteins
PubMed: 28598714
DOI: 10.1080/21688370.2017.1331155 -
STAR Protocols Dec 2021flight muscles are highly enriched with mitochondria and have emerged as a powerful genetic system for studying how oxidative phosphorylation (OXPHOS) complexes are...
flight muscles are highly enriched with mitochondria and have emerged as a powerful genetic system for studying how oxidative phosphorylation (OXPHOS) complexes are assembled. Here, we describe a series of protocols for analyzing the integrity of OXPHOS complexes in via blue native polyacrylamide gel electrophoresis (BN PAGE). We have also included protocols for the additional steps that are typically performed after OXPHOS complexes are separated by BN PAGE, such as Coomassie staining, silver staining, and in-gel OXPHOS activities. For complete details on the use and execution of this protocol, please refer to Murari et al. (2020).
Topics: Animals; Drosophila; Drosophila Proteins; Female; Flight, Animal; Male; Muscle Proteins; Muscles; Native Polyacrylamide Gel Electrophoresis; Oxidative Phosphorylation
PubMed: 34977670
DOI: 10.1016/j.xpro.2021.101021 -
BMC Developmental Biology Jan 2013Group B Sox domain transcription factors play important roles in metazoan central nervous system development. They are, however, difficult to study as mutations often...
BACKGROUND
Group B Sox domain transcription factors play important roles in metazoan central nervous system development. They are, however, difficult to study as mutations often have pleiotropic effects and other Sox family members can mask phenotypes due to functional compensation. In Drosophila melanogaster, the Sox gene Dichaete is dynamically expressed in the embryonic CNS, where it is known to have functional roles in neuroblasts and the ventral midline. In this study, we use inducible dominant negative proteins in combination with ChIP, immunohistochemistry and genome-wide expression profiling to further dissect the role of Dichaete in these two tissues.
RESULTS
We generated two dominant negative Dichaete constructs, one lacking a DNA binding domain and the other fused to the Engrailed transcriptional repressor domain. We expressed these tissue-specifically in the midline and in neuroblasts using the UAS/GAL4 system, validating their use at the phenotypic level and with known target genes. Using ChIP and immunohistochemistry, we identified two new likely direct Dichaete target genes, commisureless in the midline and asense in the neuroectoderm. We performed genome-wide expression profiling in stage 8-9 embryos, identifying almost a thousand potential tissue-specific Dichaete targets, with half of these genes showing evidence of Dichaete binding in vivo. These include a number of genes with known roles in CNS development, including several components of the Notch, Wnt and EGFR signalling pathways.
CONCLUSIONS
As well as identifying commisureless as a target, our data indicate that Dichaete helps establish its expression during early midline development but has less effect on its established later expression, highlighting Dichaete action on tissue specific enhancers. An analysis of the broader range of candidate Dichaete targets indicates that Dichaete plays diverse roles in CNS development, with the 500 or so Dichaete-bound putative targets including a number of transcription factors, signalling pathway components and terminal differentiation genes. In the early neurectoderm we implicate Dichaete in the lateral inhibition pathway and show that Dichaete acts to repress the proneural gene asense. Our analysis also reveals that dominant negatives cause off-target effects, highlighting the need to use other experimental data for validating findings from dominant negative studies.
Topics: Animals; Binding Sites; Body Patterning; Central Nervous System; Drosophila Proteins; Drosophila melanogaster; Gene Expression Regulation, Developmental; Mutation; SOX Transcription Factors
PubMed: 23289785
DOI: 10.1186/1471-213X-13-1 -
Biology of the Cell Oct 2002The formation of a belt-like junctional complex separating the apical from the lateral domain is an essential step in the differentiation of epithelial cells. Thus... (Review)
Review
The formation of a belt-like junctional complex separating the apical from the lateral domain is an essential step in the differentiation of epithelial cells. Thus protein complexes regulating this event are of first importance for the development of cell polarity and physiological functions of epithelial tissues. In Drosophila, the discovery of a gene, crb, controlling the coalescence of the spots of zonula adherens (ZA) into a adhesive ring around the cells was a major step. We know now that Crumbs, the product of crb is an apical transmembrane protein conserved in mammals and that it interacts by its cytoplasmic domain with two cortical modular proteins, Stardust (Sdt) and Discs lost (Dlt) that are also essential for the correct assembly of the ZA. These two proteins are also conserved in mammals and it is most likely that the Crumbs complex plays a similar role in very different species. Recently, we have shown that Crumbs interacts with the cortical cytoskeleton made of DMoesin and beta heavy-Spectrin and this connection could explain in part the role of Crumbs in building the ZA. Future work will help to understand several aspects of the Crumbs complex that are still unknown, like the role of the large extracellular domain or the precise function of Sdt and Dlt in the building of the ZA. Finding an answer to these questions will help to find new therapies for Retinitis pigmentosa and other retina degeneration in which CRB1, the human homologue of crb, has been involved.
Topics: Animals; Cell Polarity; Cytoskeleton; Drosophila Proteins; Epithelial Cells; Eye Proteins; Humans; Intercellular Junctions; Mammals; Membrane Proteins; Nerve Tissue Proteins
PubMed: 12500938
DOI: 10.1016/s0248-4900(02)00004-7 -
Nature Reviews. Molecular Cell Biology May 2022
Topics: Animals; Drosophila Proteins; Drosophila melanogaster; Oocytes; Protein Biosynthesis; RNA-Binding Proteins
PubMed: 35338358
DOI: 10.1038/s41580-022-00477-8 -
Nature Reviews. Cancer Mar 2007Intense research over the past four years has led to the discovery and characterization of a novel signalling network, known as the Salvador-Warts-Hippo (SWH) pathway,... (Review)
Review
Intense research over the past four years has led to the discovery and characterization of a novel signalling network, known as the Salvador-Warts-Hippo (SWH) pathway, involved in tissue growth control in Drosophila melanogaster. At present, eleven proteins have been implicated as members of this pathway, and several downstream effector genes have been characterized. The importance of this pathway is emphasized by its evolutionary conservation, and by increasing evidence that its deregulation occurs in human tumours. Here, we review the main findings from Drosophila and the implications that these have for tumorigenesis in mammals.
Topics: Animals; Cell Cycle Proteins; Drosophila Proteins; Humans; Intracellular Signaling Peptides and Proteins; Protein Kinases; Protein Serine-Threonine Kinases; Signal Transduction
PubMed: 17318211
DOI: 10.1038/nrc2070 -
Current Biology : CB Mar 2021The sex-determining genes Double Sex and Fruitless are expressed in sexually differentiated neurons of the Drosophila brain. A tiny cluster of neurons, aDN cells, serves...
The sex-determining genes Double Sex and Fruitless are expressed in sexually differentiated neurons of the Drosophila brain. A tiny cluster of neurons, aDN cells, serves as a key circuit switch with sexually dimorphic properties: those of female flies respond to visual signals in males, while those of male flies respond to smell and humidity in females, supporting effective courtship and communal egg laying behaviors, respectively.
Topics: Animals; Brain; Courtship; Drosophila; Drosophila Proteins; Female; Male; Sex Characteristics; Transcription Factors
PubMed: 33756143
DOI: 10.1016/j.cub.2021.02.026 -
PloS One 2020Genetically encoded pH indicators (GEpHI) have emerged as important tools for investigating intracellular pH (pHi) dynamics in Drosophila. However, most of the...
Genetically encoded pH indicators (GEpHI) have emerged as important tools for investigating intracellular pH (pHi) dynamics in Drosophila. However, most of the indicators are based on the Gal4/UAS binary expression system. Here, we report the generation of a ubiquitously-expressed GEpHI. The fusion protein of super ecliptic pHluorin and FusionRed was cloned under the tubulin promoter (tpHusion) to drive it independently of the Gal4/UAS system. The function of tpHusion was validated in various tissues from different developmental stages of Drosophila. Differences in pHi were also indicated correctly in fixed tissues. Finally, we describe the use of tpHusion for comparative analysis of pHi in manipulated clones and the surrounding cells in epithelial tissues. Our findings establish tpHusion as a robust tool for studying pHi in Drosophila.
Topics: Animals; Animals, Genetically Modified; Drosophila Proteins; Drosophila melanogaster; Embryo, Nonmammalian; Epithelium; Gene Expression Regulation, Developmental; Hydrogen-Ion Concentration; Transcription Factors
PubMed: 32059043
DOI: 10.1371/journal.pone.0228995