The Oscillating miRNA 959-964 cluster impacts Drosophila feeding time and other circadian outputs.
Ontology highlight
ABSTRACT:
ORGANISM(S): Drosophila melanogaster
SUBMITTER: Stefan Pescatore
PROVIDER: E-GEOD-40981 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Similar Datasets
Project description:Using high throughput sequencing of Drosophila head RNA, a small set of miRNAs that undergo robust circadian oscillations in levels were discovered. We concentrated on a cluster of six miRNAs, mir-959-964, all of which peak at about ZT12 or lights-off. The data indicate that the cluster pri-miRNA is transcribed under bona fide circadian transcriptional control and that all 6 mature miRNAs have short half-lives, a requirement for oscillating. Manipulation of food intake dramatically affects the levels and timing of cluster miRNA transcription with no more than minor effects on the core circadian oscillator. This indicates that the central clock regulates feeding, which in turn regulates proper levels and cycling of the cluster miRNAs. Viable Gal4 knock-in as well as cluster knock-out and over-expression strains were used to localize cluster miRNA expression as well as explore their functions. The adult head fat body is a major site of expression, and feeding behavior, innate immunity, metabolism, and perhaps stress responses are under cluster miRNA regulation. The feeding behavior results indicate that there is a feedback circuit between feeding time and cluster miRNA function as well as a surprising role of post-transcriptional regulation in these behaviors and physiology. Six samples of small RNA libraries (RNA size 19 to 29 nucleotides long) were prepared from Drosophila heads, each collected at one circadian time point during a light-dark cycle (ZT0, ZT4, ZT8, ZT12, ZT16, ZT20).
2012-11-02 | E-GEOD-40943 | biostudies-arrayexpress
Project description:Using high throughput sequencing of Drosophila head RNA, a small set of miRNAs that undergo robust circadian oscillations in levels were discovered. We concentrated on a cluster of six miRNAs, mir-959-964, all of which peak at about ZT12 or lights-off. The data indicate that the cluster pri-miRNA is transcribed under bona fide circadian transcriptional control and that all 6 mature miRNAs have short half-lives, a requirement for oscillating. Manipulation of food intake dramatically affects the levels and timing of cluster miRNA transcription with no more than minor effects on the core circadian oscillator. This indicates that the central clock regulates feeding, which in turn regulates proper levels and cycling of the cluster miRNAs. Viable Gal4 knock-in as well as cluster knock-out and over-expression strains were used to localize cluster miRNA expression as well as explore their functions. The adult head fat body is a major site of expression, and feeding behavior, innate immunity, metabolism, and perhaps stress responses are under cluster miRNA regulation. The feeding behavior results indicate that there is a feedback circuit between feeding time and cluster miRNA function as well as a surprising role of post-transcriptional regulation in these behaviors and physiology. To address possible functions of the cluster miRNAs, the knock-out strain (mir959-952-KO) as well as a cluster over-expression strain (UAS-cluster, tim-gal4) was assayed for mRNA changes relative to their WT counterparts on Affymetrix expression arrays. The strategy was based on the observation that miRNAs often cause a decrease in the steady-state levels of their target mRNAs (Guo et al., 2010; Lim et al., 2005). Because of the circadian regulation and the possibility of non-fat body expression, the over-expression strain was generated with the broad circadian driver tim-gal4 rather than a fat body driver. To accommodate the possibility that important mRNA changes only appear at certain circadian times, RNA was assayed from heads collected at two different times, ZT4 and ZT16. Analyzed expression data was assayed from total RNA from Drosophila heads in two sample groups consisting of four samples (with eight samples in total): knock-out strain at ZT4 and ZT16, and its respective wildtype control at ZT4 and ZT16; cluster over-expression strain at ZT4 and ZT16 and its respective wildtype control at ZT4 and ZT16.
2012-11-02 | E-GEOD-40894 | biostudies-arrayexpress
Project description:The Oscillating miRNA 959-964 cluster impacts Drosophila feeding time and other circadian outputs [expression].
| PRJNA175269 | ENA
Project description:The Oscillating miRNA 959-964 cluster impacts Drosophila feeding time and other circadian outputs [miRNA-seq].
| PRJNA175420 | ENA
Project description:MicroRNAs are a wide class of ~22 nt non-coding RNAs of metazoans capable of inhibiting target mRNAs translation by binding to partially complementary sites in their 3âUTRs. Due to their regulatory potential, miRNAs are implicated in functioning of a broad range of biological pathways and processes. Here we investigate the functions of the miR-959-964 cluster expressed predominantly in testes of Drosophila melanogaster. The deletion of miR-959-964 resulted in male sterility due to the disturbance of the spermatid individualization process. Analysis of the transcriptome by microarray followed by luciferase reporter assay revealed didum, for, fdl and CG10512 as the targets of miR-959-964. Moreover, the deletion of miR-959-964 is accompanied by a decreased the expression of genes responsible for microtubule-based movement and spermatid differentiation. Thus, we suggest that miR-959-964 can control the process of spermatid individualization by direct and indirect modulating the expression of different components of the individualization process. In addition, we have shown that in comparison to other miRNAs, the rate of evolution of the testis-specific miR-959-964 cluster is unusually high, indicating its possible involvement in speciation via reproductive isolation.
2015-10-29 | E-MTAB-3214 | biostudies-arrayexpress
Project description:Aβ peptides derived from the amyloid precursor protein (APP) have been strongly implicated in the pathogenesis of Alzheimer’s disease. However, the normal function of APP and the importance of that role in neurodegenerative disease is less clear. We recovered the Drosophila ortholog of APP, Appl, in an unbiased forward genetic screen for neurodegeneration mutants. We performed comprehensive single cell transcriptional and proteomic studies of Appl mutant flies to investigate Appl function in the aging brain.
2023-10-04 | PXD041862 | Pride
Project description:Here we describe a bottom-up approach to analyse an enriched membrane fraction from Drosophila melanogaster heads using multidimensional liquid chromatography (LC) coupled with tandem-mass spectrometry (MS/MS) that relies on a complete solubilisation and digestion of proteins. An enriched membrane fraction was prepared using equilibrium density centrifugation on a discontinual sucrose gradient, followed by solubilisation using FASP method, tryptic and consequential chymotryptic digestion of proteins. Peptides were separated by reversed-phase (RP) LC at high pH in the first dimension and acidic RP-LC coupled directly to Orbitrap Velos Pro mass spectrometer.
2015-07-27 | PXD001712 | Pride
Project description:We sequenced mRNA extracted from heads of a D. melanogaster population that was sedated with a stream of ethanol saturated vapor, 30 minutes before RNA extraction; and from an age-matched untreated control group. Differential gene expression between the two groups was calculated and reported. Examination of mRNA levels in heads of D. melanogaster adult females after ethanol exposure was performed using next generation sequencing (NGS) technology.
2016-02-11 | E-GEOD-77792 | biostudies-arrayexpress
Project description:Broadly expressed transcriptions factors (TFs) control tissue-specific programs of gene expression through interactions with local TF networks. Prime examples are the circadian clock TFs CLOCK (CLK) and CYCLE (CYC or BMAL1): while they control a core transcriptional circuit throughout animal bodies, downstream clock target genes and circadian physiology are tissue-specific. Here, we use ChIP-seq to determine the regulatory targets of Drosophila CLK and CYC, which we epitope-tagged by homologous recombination. Both TFs have distinct binding sites in heads versus bodies, suggesting that they directly control tissue-specific downstream target genes. Analysis of these context-specific binding sites revealed distinct sequence motifs for putative clock partner factors, including a motif for the GATA factor SERPENT (SRP). SRP indeed synergistically enhances CLK/CYC-mediated activity of a cis-regulatory region bound by CLK/CYC specifically in bodies. These results reveal how universal clock circuits can generate tissue-specific outputs and demonstrate an approach to dissect regulatory interactions more generally. We sequenced ChIP and input samples, as well as M-bM-^@M-^\mockM-bM-^@M-^] samples for which we performed ChIP with the V5 antibody from wildtype w- flies (not carrying the V5 tag) for two independent biological replicates each, summing to 24 libraries in total.
2013-06-01 | E-GEOD-40467 | biostudies-arrayexpress