Genes encoding the KDM5 family of transcriptional regulators are disrupted in individuals with intellectual disability (ID). To understand the link between KDM5 and ID, we characterized five Drosophila strains harboring missense alleles analogous to those observed in patients. These alleles disrupted neuroanatomical development, cognition and other behaviors, and displayed a transcriptional signature characterized by the downregulation of many ribosomal protein genes. A similar transcriptional profile was observed in KDM5C knockout iPSC-induced human glutamatergic neurons, suggesting an evolutionarily conserved role for KDM5 proteins in regulating this class of gene. In Drosophila, reducing KDM5 changed neuronal ribosome composition, lowered the translation efficiency of mRNAs required for mitochondrial function, and altered mitochondrial metabolism. These data highlight the cellular consequences of altered KDM5-regulated transcriptional programs that could contribute to cognitive and behavioral phenotypes. Moreover, they suggest that KDM5 may be part of a broader network of proteins that influence cognition by regulating protein synthesis.

• MeSH
• aktivace transkripce MeSH
• Drosophila melanogaster genetika   metabolismus   MeSH
• Drosophila genetika   metabolismus   MeSH
• histondemethylasy metabolismus   genetika   MeSH
• lidé MeSH
• mentální retardace genetika   metabolismus   MeSH
• mitochondrie metabolismus   genetika   MeSH
• neurony * metabolismus   MeSH
• proteiny Drosophily * genetika   metabolismus   MeSH
• proteosyntéza MeSH
• ribozomální proteiny * genetika   metabolismus   MeSH
• ribozomy metabolismus   genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• histondemethylasy MeSH
• KDM5C protein, human MeSH Prohlížeč
• Lid protein, Drosophila MeSH Prohlížeč
• proteiny Drosophily * MeSH
• ribozomální proteiny * MeSH

BACKGROUND: The metabolically demanding nature of immune response requires nutrients to be preferentially directed towards the immune system at the expense of peripheral tissues. We study the mechanisms by which this metabolic reprograming occurs using the parasitoid infection of Drosophila larvae. To overcome such an immune challenge hemocytes differentiate into lamellocytes, which encapsulate and melanize the parasitoid egg. Hemocytes acquire the energy for this process by expressing JAK/STAT ligands upd2 and upd3, which activates JAK/STAT signaling in muscles and redirects carbohydrates away from muscles in favor of immune cells. METHODS: Immune response of Drosophila larvae was induced by parasitoid wasp infestation. Carbohydrate levels, larval locomotion and gene expression of key proteins were compared between control and infected animals. Efficacy of lamellocyte production and resistance to wasp infection was observed for RNAi and mutant animals. RESULTS: Absence of upd/JAK/STAT signaling leads to an impaired immune response and increased mortality. We demonstrate how JAK/STAT signaling in muscles leads to suppression of insulin signaling through activation of ImpL2, the inhibitor of Drosophila insulin like peptides. CONCLUSIONS: Our findings reveal cross-talk between immune cells and muscles mediates a metabolic shift, redirecting carbohydrates towards immune cells. We emphasize the crucial function of muscles during immune response and show the benefits of insulin resistance as an adaptive mechanism that is necessary for survival.

• Klíčová slova
• Drosophila, Immunity, Insulin resistance, JAK/STAT signaling, Metabolism,
• MeSH
• Drosophila genetika   MeSH
• imunita MeSH
• inzulinová rezistence * MeSH
• Janus kinasy metabolismus   MeSH
• larva metabolismus   MeSH
• proteiny Drosophily * metabolismus   MeSH
• proteiny vázající insulinu podobné růstové faktory metabolismus   MeSH
• sacharidy MeSH
• sršňovití * metabolismus   MeSH
• svaly MeSH
• transkripční faktory STAT metabolismus   MeSH
• transkripční faktory metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• ImpL2 protein, Drosophila MeSH Prohlížeč
• Janus kinasy MeSH
• proteiny Drosophily * MeSH
• proteiny vázající insulinu podobné růstové faktory MeSH
• sacharidy MeSH
• transkripční faktory STAT MeSH
• transkripční faktory MeSH

Genetic screens have been used extensively to probe interactions between nuclear genes and their impact on phenotypes. Probing interactions between mitochondrial genes and their phenotypic outcome, however, has not been possible due to a lack of tools to map the responsible polymorphisms. Here, using a toolkit we previously established in Drosophila, we isolate over 300 recombinant mitochondrial genomes and map a naturally occurring polymorphism at the cytochrome c oxidase III residue 109 (CoIII109) that fully rescues the lethality and other defects associated with a point mutation in cytochrome c oxidase I (CoIT300I). Through lipidomics profiling, biochemical assays and phenotypic analyses, we show that the CoIII109 polymorphism modulates cardiolipin binding to prevent complex IV instability caused by the CoIT300I mutation. This study demonstrates the feasibility of genetic interaction screens in animal mitochondrial DNA. It unwraps the complex intra-genomic interplays underlying disorders linked to mitochondrial DNA and how they influence disease expression.

• MeSH
• Drosophila genetika   MeSH
• kardiolipiny * genetika   metabolismus   MeSH
• mitochondriální DNA * genetika   metabolismus   MeSH
• mitochondrie genetika   metabolismus   MeSH
• respirační komplex IV metabolismus   MeSH
• umělé letální mutace MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• kardiolipiny * MeSH
• mitochondriální DNA * MeSH
• respirační komplex IV MeSH

Target-AID, BE3, and ABE7.10 base editors fused to the catalytically modified Cas9 and xCas9(3.7) were tested for germline editing of the fruit fly Drosophila melanogaster. We developed a guide RNA-expressing construct, white-4gRNA, targeting splice sites in the white gene, an X-chromosome located gene. Using white-4gRNA flies and transgenic lines expressing Target-AID, BE3, and ABE7.10 base editors, we tested the efficiency of stable germline gene editing at three different temperatures. Classical Cas9 generating insertions/deletions by non-homologous end joining served as a reference. Our data indicate that gene editing is most efficient at 28°C, the highest temperature suitable for fruit flies. Finally, we created a new allele of the core circadian clock gene timeless using Target-AID. This base edited mutant allele timSS308-9FL had a disrupted circadian clock with a period of ∼29 h. The white-4gRNA expressing fly can be used to test new generations of base editors for future applications in Drosophila.

• MeSH
• adenin MeSH
• CRISPR-Cas systémy * genetika   MeSH
• cytosin MeSH
• Drosophila melanogaster genetika   MeSH
• Drosophila genetika   MeSH
• editace genu * MeSH
• vodící RNA, systémy CRISPR-Cas MeSH
• zárodečné buňky MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adenin MeSH
• cytosin MeSH
• vodící RNA, systémy CRISPR-Cas MeSH

Telomeres in Drosophila melanogaster, which have inspired a large part of Sergio Pimpinelli work, are similar to those of other eukaryotes in terms of their function. Yet, their length maintenance relies on the transposition of the specialized retrotransposons Het-A, TART, and TAHRE, rather than on the activity of the enzyme telomerase as it occurs in most other eukaryotic organisms. The length of the telomeres in Drosophila thus depends on the number of copies of these transposable elements. Our previous work has led to the isolation of a dominant mutation, Tel1, that caused a several-fold elongation of telomeres. In this study, we molecularly identified the Tel1 mutation by a combination of transposon-induced, site-specific recombination and next-generation sequencing. Recombination located Tel1 to a 15 kb region in 92A. Comparison of the DNA sequence in this region with the Drosophila Genetic Reference Panel of wild-type genomic sequences delimited Tel1 to a 3 bp deletion inside intron 8 of Ino80. Furthermore, CRISPR/Cas9-induced deletions surrounding the same region exhibited the Tel1 telomere phenotype, confirming a strict requirement of this intron 8 gene sequence for a proper regulation of Drosophila telomere length.

• Klíčová slova
• Drosophila melanogaster, next-generation sequencing, telomere, transposon-induced recombination,
• MeSH
• Drosophila melanogaster * genetika   MeSH
• Drosophila * genetika   MeSH
• genové produkty gag genetika   MeSH
• mutace genetika   MeSH
• telomery genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• genové produkty gag MeSH

In animals and plants, Dicer enzymes collaborate with double-stranded RNA-binding domain (dsRBD) proteins to convert precursor-microRNAs (pre-miRNAs) into miRNA duplexes. We report six cryo-EM structures of Drosophila Dicer-1 that show how Dicer-1 and its partner Loqs‑PB cooperate (1) before binding pre-miRNA, (2) after binding and in a catalytically competent state, (3) after nicking one arm of the pre-miRNA, and (4) following complete dicing and initial product release. Our reconstructions suggest that pre-miRNA binds a rare, open conformation of the Dicer‑1⋅Loqs‑PB heterodimer. The Dicer-1 dsRBD and three Loqs‑PB dsRBDs form a tight belt around the pre-miRNA, distorting the RNA helix to place the scissile phosphodiester bonds in the RNase III active sites. Pre-miRNA cleavage shifts the dsRBDs and partially closes Dicer-1, which may promote product release. Our data suggest a model for how the Dicer‑1⋅Loqs‑PB complex affects a complete cycle of pre-miRNA recognition, stepwise endonuclease cleavage, and product release.

• Klíčová slova
• Dcr-1, Dicer, Dicer-partner proteins, Loqs-PB, Loquacious, RNase III, cryo-EM, dsRBD, isomiR, miRNA, microRNA,
• MeSH
• Drosophila genetika   MeSH
• mikro RNA * genetika   metabolismus   MeSH
• proteiny Drosophily * genetika   metabolismus   MeSH
• proteiny vázající RNA metabolismus   MeSH
• ribonukleasa III genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, N.I.H., Intramural MeSH
• Názvy látek
• mikro RNA * MeSH
• proteiny Drosophily * MeSH
• proteiny vázající RNA MeSH
• ribonukleasa III MeSH

The adenosine deaminase acting on RNA (ADAR) enzymes are essential for neuronal function and innate immune control. ADAR1 RNA editing prevents aberrant activation of antiviral dsRNA sensors through editing of long, double-stranded RNAs (dsRNAs). In this review, we focus on the ADAR2 proteins involved in the efficient, highly site-specific RNA editing to recode open reading frames first discovered in the GRIA2 transcript encoding the key GLUA2 subunit of AMPA receptors; ADAR1 proteins also edit many of these sites. We summarize the history of ADAR2 protein research and give an up-to-date review of ADAR2 structural studies, human ADARBI (ADAR2) mutants causing severe infant seizures, and mouse disease models. Structural studies on ADARs and their RNA substrates facilitate current efforts to develop ADAR RNA editing gene therapy to edit disease-causing single nucleotide polymorphisms (SNPs). Artificial ADAR guide RNAs are being developed to retarget ADAR RNA editing to new target transcripts in order to correct SNP mutations in them at the RNA level. Site-specific RNA editing has been expanded to recode hundreds of sites in CNS transcripts in Drosophila and cephalopods. In Drosophila and C. elegans, ADAR RNA editing also suppresses responses to self dsRNA.

• Klíčová slova
• ADAR, ADARB1, dsRNA, neurons, recoding RNA editing,
• MeSH
• adenosindeaminasa * metabolismus   MeSH
• AMPA receptory genetika   metabolismus   MeSH
• antivirové látky MeSH
• Caenorhabditis elegans genetika   MeSH
• Drosophila genetika   MeSH
• dvouvláknová RNA genetika   MeSH
• genetická terapie MeSH
• lidé MeSH
• myši MeSH
• proteiny vázající RNA genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• ADARB1 protein, human MeSH Prohlížeč
• adenosindeaminasa * MeSH
• AMPA receptory MeSH
• antivirové látky MeSH
• dvouvláknová RNA MeSH
• proteiny vázající RNA MeSH

The rather recent development of genetically encoded metabolite sensors has changed the way we can study metabolism in living cells, ex vivo tissues, and in vivo immensely. In recent years, these sensors have also been adapted for use in Drosophila tissues. Here, we describe a standard protocol to image such sensors in ex vivo Drosophila larval brains using the glucose sensor FLII12Pglu-700μδ6. The protocol, however, can be adapted for the use of other sensors, tissues, and can even be used in vivo.

• Klíčová slova
• Carbohydrate transport, FRET, Genetically encoded metabolite sensors, Live imaging, Metabolism,
• MeSH
• biosenzitivní techniky * metody   MeSH
• Drosophila genetika   MeSH
• rezonanční přenos fluorescenční energie * metody   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The ER membrane protein complex (EMC) is required for the biogenesis of a subset of tail anchored (TA) and polytopic membrane proteins, including Rhodopsin-1 (Rh1) and the TRP channel. To understand the physiological implications of EMC-dependent membrane protein biogenesis, we perform a bioinformatic identification of Drosophila TA proteins. From 254 predicted TA proteins, screening in larval eye discs identified two proteins that require EMC for their biogenesis: fan and Xport-A. Fan is required for male fertility in Drosophila and we show that EMC is also required for this process. Xport-A is essential for the biogenesis of both Rh1 and TRP, raising the possibility that disruption of Rh1 and TRP biogenesis in EMC mutants is secondary to the Xport-A defect. We show that EMC is required for Xport-A TMD membrane insertion and that EMC-independent Xport-A mutants rescue Rh1 and TRP biogenesis in EMC mutants. Finally, our work also reveals a role for Xport-A in a glycosylation-dependent triage mechanism during Rh1 biogenesis in the endoplasmic reticulum.

• Klíčová slova
• ER membrane protein complex, Rh1, TRP, Xport-A, tail anchored proteins,
• MeSH
• Drosophila genetika   metabolismus   MeSH
• endoplazmatické retikulum metabolismus   MeSH
• membránové proteiny genetika   metabolismus   MeSH
• molekulární chaperony * genetika   metabolismus   MeSH
• proteiny Drosophily * genetika   metabolismus   MeSH
• represorové proteiny * genetika   metabolismus   MeSH
• rodopsin * genetika   MeSH
• transkripční faktory bHLH * genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• emc protein, Drosophila MeSH Prohlížeč
• membránové proteiny MeSH
• molekulární chaperony * MeSH
• proteiny Drosophily * MeSH
• represorové proteiny * MeSH
• rodopsin * MeSH
• transkripční faktory bHLH * MeSH
• Xport-A protein, Drosophila MeSH Prohlížeč

Molecular identification is increasingly used to speed up biodiversity surveys and laboratory experiments. However, many groups of organisms cannot be reliably identified using standard databases such as GenBank or BOLD due to lack of sequenced voucher specimens identified by experts. Sometimes a large number of sequences are available, but with too many errors to allow identification. Here, we address this problem for parasitoids of Drosophila by introducing a curated open-access molecular reference database, DROP (Drosophila parasitoids). Identifying Drosophila parasitoids is challenging and poses a major impediment to realize the full potential of this model system in studies ranging from molecular mechanisms to food webs, and in biological control of Drosophila suzukii. In DROP, genetic data are linked to voucher specimens and, where possible, the voucher specimens are identified by taxonomists and vetted through direct comparison with primary type material. To initiate DROP, we curated 154 laboratory strains, 856 vouchers, 554 DNA sequences, 16 genomes, 14 transcriptomes, and six proteomes drawn from a total of 183 operational taxonomic units (OTUs): 114 described Drosophila parasitoid species and 69 provisional species. We found species richness of Drosophila parasitoids to be heavily underestimated and provide an updated taxonomic catalogue for the community. DROP offers accurate molecular identification and improves cross-referencing between individual studies that we hope will catalyse research on this diverse and fascinating model system. Our effort should also serve as an example for researchers facing similar molecular identification problems in other groups of organisms.

• Klíčová slova
• DNA sequences, biodiversity, biological control, genomes, integrative taxonomy, molecular diagnostics,
• MeSH
• biodiverzita * MeSH
• Drosophila * genetika   MeSH
• potravní řetězec MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH