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Archives of Insect Biochemistry and... Mar 2020Insect metamorphosis is regulated by two main hormones: ecdysone (20E), which promotes molting, and juvenile hormone (JH), which inhibits adult morphogenesis. The... (Review)
Review
Insect metamorphosis is regulated by two main hormones: ecdysone (20E), which promotes molting, and juvenile hormone (JH), which inhibits adult morphogenesis. The transduction mechanisms for the respective hormonal signals include the transcription factors Krüppel homolog 1 (Kr-h1) and E93, which are JH- and 20E-dependent, respectively. Kr-h1 is the main effector of the antimetamorphic action of JH, while E93 is a key promoter of metamorphosis. The ancestral regulatory axis of metamorphosis, which operates in insects with hemimetabolan (gradual) metamorphosis and is known as the MEKRE93 pathway, is based on Kr-h1 repression of E93. In the last juvenile stage, when the production of JH dramatically decreases, Kr-h1 expression is almost completely interrupted, E93 becomes upregulated and metamorphosis proceeds. The holometabolan (complete) metamorphosis mode of development includes the peculiar pupal stage, a sort of intermediate between the final larval instar and the adult stage. In holometabolan species, Broad-Complex (BR-C) transcription factors determine the pupal stage and E93 stimulates the expression of BR-C in the prepupa. The MEKRE93 pathway is conserved in holometabolan insects, which have added the E93/BR-C interaction loop to the ancestral (hemimetabolan) pathway during the evolution from hemimetaboly to holometaboly.
Topics: Animals; Drosophila Proteins; Gene Expression Regulation, Developmental; Kruppel-Like Transcription Factors; Metamorphosis, Biological; Transcription Factors
PubMed: 31385626
DOI: 10.1002/arch.21609 -
Physiological Reports Feb 2018Cardiolipin (CL) is a mitochondrial phospholipid that helps maintain normal structure of the inner mitochondrial membrane and stabilize the protein complexes of the...
Cardiolipin (CL) is a mitochondrial phospholipid that helps maintain normal structure of the inner mitochondrial membrane and stabilize the protein complexes of the electron transport chain to promote efficient ATP synthesis. Tafazzin, an acyl-transferase, is required for synthesis of the mature form of CL. Mutations in the tafazzin (TAZ) gene are associated with a human disorder known as Barth syndrome. Symptoms of Barth syndrome often include muscle weakness and exercise intolerance. Previous work demonstrates that Drosophila Taz mutants exhibit motor weakness, as measured by reduced flying and climbing abilities. However, Drosophila TAZ mutants' baseline endurance or response to endurance exercise training has not been assessed. Here, we find that TAZ mutants have reduced endurance and do not improve following a stereotypical exercise training paradigm, indicating that loss of TAZ function leads to exercise intolerance in Drosophila. Although cardiac phenotypes are observed in human Barth syndrome patients, TAZ mutants had normal resistance to cardiac pacing. In the future, endurance may be a useful screening tool to identify additional genetic modifiers of tafazzin.
Topics: 1-Acylglycerophosphocholine O-Acyltransferase; Animals; Drosophila; Drosophila Proteins; Motor Neurons; Muscle, Skeletal; Mutation; Physical Conditioning, Animal
PubMed: 29405656
DOI: 10.14814/phy2.13604 -
International Journal of Molecular... Mar 2023Lipid homeostasis is essential for insect growth and development. The complex of proteins associated with Set 1 (COMPASS)-catalyzed Histone 3 lysine 4 trimethylation...
Lipid homeostasis is essential for insect growth and development. The complex of proteins associated with Set 1 (COMPASS)-catalyzed Histone 3 lysine 4 trimethylation (H3K4me3) epigenetically activates gene transcription and is involved in various biological processes, but the role and molecular mechanism of H3K4me3 modification in lipid homeostasis remains largely unknown. In the present study, we showed in that fat body-specific knockdown of () as one of the COMPASS complex components caused a decrease in lipid droplet (LD) size and triglyceride (TG) levels. Mechanistically, Wds-mediated H3K4me3 modification in the fat body targeted several lipogenic genes involved in lipid synthesis and the gene associated with lipid transport to promote their expressions; the transcription factor heat shock factor (Hsf) could interact with Wds to modulate H3K4me3 modification within the promoters of these targets; and fat body-specific knockdown of phenocopied the effects of knockdown on lipid homeostasis in the fat body. Moreover, fat body-specific knockdown of or reduced high-fat diet (HFD)-induced oversized LDs and high TG levels. Altogether, our study reveals that Wds-mediated H3K4me3 modification is required for lipid homeostasis during development and provides novel insights into the epigenetic regulation of insect lipid metabolism.
Topics: Animals; Histones; Drosophila; Epigenesis, Genetic; Drosophila Proteins; Lipids
PubMed: 37047100
DOI: 10.3390/ijms24076125 -
Cells May 2020The cell line 1182-4, which constitutively lacks centrioles, was established many years ago from haploid embryos laid by females homozygous for the mutation. This was... (Review)
Review
The cell line 1182-4, which constitutively lacks centrioles, was established many years ago from haploid embryos laid by females homozygous for the mutation. This was the first clear example of animal cells regularly dividing in the absence of this organelle. However, the cause of the acentriolar nature of the 1182-4 cell line remained unclear and could not be clearly assigned to a particular genetic event. Here, we detail historically the longstanding mystery of the lack of centrioles in this cell line. Recent advances, such as the characterization of the gene and the genomic analysis of 1182-4 cells, allow now a better understanding of the physiology of these cells. By combining these new data, we propose three reasonable hypotheses of the genesis of this remarkable phenotype.
Topics: Animals; Cell Line; Centrioles; Drosophila Proteins; Drosophila melanogaster; Genome, Insect; Models, Biological
PubMed: 32456186
DOI: 10.3390/cells9051300 -
Methods in Molecular Biology (Clifton,... 2019Molecular dynamics (MD) simulations enable in silico investigations of the dynamic behavior of proteins and protein complexes. Here, we describe MD simulations of the...
Molecular dynamics (MD) simulations enable in silico investigations of the dynamic behavior of proteins and protein complexes. Here, we describe MD simulations of the SNARE complex and its interactions with the neuronal protein complexin. Complexin is an effector of neuronal secretion that inhibits spontaneous fusion and is thought to clamp the fusion process via the interactions with the SNARE complex. We describe MD simulations of the SNARE complex alone and bound to complexin. The MD simulations under external forces imitating the repulsion between lipid bilayers enabled us to investigate unraveling and assembly of the SNARE complex.
Topics: Adaptor Proteins, Vesicular Transport; Crystallography, X-Ray; Drosophila Proteins; Lipid Bilayers; Models, Biological; Molecular Dynamics Simulation; Protein Binding; SNARE Proteins; Sequence Homology, Amino Acid
PubMed: 30317495
DOI: 10.1007/978-1-4939-8760-3_1 -
Uspekhi Fiziologicheskikh Nauk 2015Elucidation of molecular mechanisms of cognitive functions is one of the major achievements in neurobiology. At most, this is due to the studies on the simple nervous... (Review)
Review
Elucidation of molecular mechanisms of cognitive functions is one of the major achievements in neurobiology. At most, this is due to the studies on the simple nervous systems, such as the CNS in Drosophila melanogaster. Many of its functional characteristics are pretty similar to higher vertebrates. Among these are: 1) evolutionary conservation of genes and molecular systems involved in the regulation of learning acquisition and memory formation; 2) presence of highly specialized and differentiated sensory, associative and motor centers; 3) utilization of similar modes of informational coding and analysis; 4) availability of major learning forms including non-associative, as well as associative learning; 5) diversity of different memories, including short-term- and protein synthesis- dependent long-term memory; 6) presence of aminergic reinforcement systems in the brain; 7) feed-back loops of circadian clocks, current organism experience and individual organism characters affecting cognitive process per se. In this review the main attention is paid to the two mostly studied Drosophila learning forms, namely to olfactory Iearning and courtship suppression conditioning (CCS). A separate consideration is given to the impacts of kynurenins and metabolite of actin remodeling signal cascade.
Topics: Animals; Brain; Drosophila; Drosophila Proteins; Memory
PubMed: 25854115
DOI: No ID Found -
Nature Communications Jun 2022How signaling proteins generate a multitude of information to organize tissue patterns is critical to understanding morphogenesis. In Drosophila, FGF produced in...
How signaling proteins generate a multitude of information to organize tissue patterns is critical to understanding morphogenesis. In Drosophila, FGF produced in wing-disc cells regulates the development of the disc-associated air-sac-primordium (ASP). Here, we show that FGF is Glycosylphosphatidylinositol-anchored to the producing cell surface and that this modification both inhibits free FGF secretion and promotes target-specific cytoneme contacts and contact-dependent FGF release. FGF-source and ASP cells extend cytonemes that present FGF and FGFR on their surfaces and reciprocally recognize each other over distance by contacting through cell-adhesion-molecule (CAM)-like FGF-FGFR binding. Contact-mediated FGF-FGFR interactions induce bidirectional responses in ASP and source cells that, in turn, polarize FGF-sending and FGF-receiving cytonemes toward each other to reinforce signaling contacts. Subsequent un-anchoring of FGFR-bound-FGF from the source membrane dissociates cytoneme contacts and delivers FGF target-specifically to ASP cytonemes for paracrine functions. Thus, GPI-anchored FGF organizes both source and recipient cells and self-regulates its cytoneme-mediated tissue-specific dispersion.
Topics: Animals; Drosophila; Drosophila Proteins; Drosophila melanogaster; Imaginal Discs; Wings, Animal
PubMed: 35710780
DOI: 10.1038/s41467-022-30417-1 -
Nucleic Acids Research Feb 2021Pre-mRNA splicing catalyzed by the spliceosome represents a critical step in the regulation of gene expression contributing to transcriptome and proteome diversity. The...
Pre-mRNA splicing catalyzed by the spliceosome represents a critical step in the regulation of gene expression contributing to transcriptome and proteome diversity. The spliceosome consists of five small nuclear ribonucleoprotein particles (snRNPs), the biogenesis of which remains only partially understood. Here we define the evolutionarily conserved protein Ecdysoneless (Ecd) as a critical regulator of U5 snRNP assembly and Prp8 stability. Combining Drosophila genetics with proteomic approaches, we demonstrate the Ecd requirement for the maintenance of adult healthspan and lifespan and identify the Sm ring protein SmD3 as a novel interaction partner of Ecd. We show that the predominant task of Ecd is to deliver Prp8 to the emerging U5 snRNPs in the cytoplasm. Ecd deficiency, on the other hand, leads to reduced Prp8 protein levels and compromised U5 snRNP biogenesis, causing loss of splicing fidelity and transcriptome integrity. Based on our findings, we propose that Ecd chaperones Prp8 to the forming U5 snRNP allowing completion of the cytoplasmic part of the U5 snRNP biogenesis pathway necessary to meet the cellular demand for functional spliceosomes.
Topics: Animals; Cell Line; Drosophila Proteins; Drosophila melanogaster; Mutation; Protein Stability; RNA Splicing; RNA Splicing Factors; Ribonucleoprotein, U5 Small Nuclear; Transcriptome
PubMed: 33444449
DOI: 10.1093/nar/gkaa1274 -
Protein Expression and Purification Jun 2019Homeodomain transcription factors (HD TFs) are a large class of evolutionarily conserved DNA binding proteins that contain a basic 60-amino acid region required for...
Homeodomain transcription factors (HD TFs) are a large class of evolutionarily conserved DNA binding proteins that contain a basic 60-amino acid region required for binding to specific DNA sites. In Drosophila melanogaster, many of these HD TFs are expressed in the early embryo and control transcription of target genes in development through their interaction with cis-regulatory modules. Previous studies where some of the Drosophila HD TFs were purified required the use of strong denaturants (i.e. 6 M urea) and multiple chromatography columns, making the downstream biochemical examination of the isolated protein difficult. To circumvent these obstacles, we have developed a streamlined expression and purification protocol to produce large yields of Drosophila HD TFs. Using the HD TFs FUSHI-TARAZU (FTZ), ANTENNAPEDIA (ANTP), ABDOMINAL-A (ABD-A), ABDOMINAL-B (ABD-B), and ULTRABITHORAX (UBX) as examples, we demonstrate that our 3-day protocol involving the overexpression of His-SUMO fusion constructs in E. coli followed by a Ni-IMAC, SUMO-tag cleavage with the SUMO protease Ulp1, and a heparin column purification produces pure, soluble protein in biological buffers around pH 7 in the absence of denaturants. Electrophoretic mobility shift assays (EMSA) confirm that the purified HD proteins are functional and nuclear magnetic resonance (NMR) spectra confirm that the purified HDs are well-folded. These purified HD TFs can be used in future biophysical experiments to structurally and biochemically characterize how and why these HD TFs bind to different DNA sequences and further probe how nucleotide differences contribute to TF-DNA specificity in the HD family.
Topics: Animals; Drosophila Proteins; Drosophila melanogaster; Homeodomain Proteins; Recombinant Fusion Proteins
PubMed: 30738927
DOI: 10.1016/j.pep.2019.02.001 -
Developmental Biology May 2016Regulated Apoptosis (Programmed Cell Death, PCD) maintains tissue homeostasis in adults, and ensures proper growth and morphogenesis of tissues during development of...
Regulated Apoptosis (Programmed Cell Death, PCD) maintains tissue homeostasis in adults, and ensures proper growth and morphogenesis of tissues during development of metazoans. Accordingly, defects in cellular processes triggering or executing apoptotic programs have been implicated in a variety of degenerative and neoplastic diseases. Here, we report the identification of DCAF12, an evolutionary conserved member of the WD40-motif repeat family of proteins, as a new regulator of apoptosis in Drosophila. We find that DCAF12 is required for Diap1 cleavage in response to pro-apoptotic signals, and is thus necessary and sufficient for RHG (Reaper, Hid, and Grim)-mediated apoptosis. Loss of DCAF12 perturbs the elimination of supernumerary or proliferation-impaired cells during development, and enhances tumor growth induced by loss of neoplastic tumor suppressors, highlighting the wide requirement for DCAF12 in PCD.
Topics: Alleles; Animals; Animals, Genetically Modified; Apoptosis; Caspases; Cell Proliferation; Crosses, Genetic; Drosophila Proteins; Drosophila melanogaster; Female; Gene Deletion; Homeostasis; Homozygote; Inhibitor of Apoptosis Proteins; Male; Metamorphosis, Biological; Morphogenesis; Mutation; Phenotype; Retina
PubMed: 26972874
DOI: 10.1016/j.ydbio.2016.03.003