Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

The hub protein Loqs connects the miRNA and endo-siRNA pathways in mosquitos

ABSTRACT: Aedes aegypti mosquitoes vector several arboviruses of global health significance, including dengue viruses and chikungunya virus. RNA interference (RNAi) plays an important role in antiviral immunity, gene regulation and protection from transposable elements. Double-stranded RNA binding proteins (dsRBPs) are important for efficient RNAi; in Drosophila functional specialization of the miRNA, endo-siRNA and exo-siRNA pathway is aided by the dsRBPs Loqs-PB, Loqs-PD and R2D2, respectively. However, this functional specialization has not been investigated in other dipterans. Characterization of the gene structure, expression pattern and interactions with other RNAi/miRNA components revealed that mosquito Loqs/R3D1 isoform -PA, but not -PB interacted readily with both AeAGO1 and AeAGO2. This interaction was mapped to a 32 a.a. region predicted to be structurally similar to the unique 22 a.a. tail of Drosophila Loqs-PD. No -PD isoforms could be detected in Ae. aegypti; analysis of other dipteran genomes demonstrated that this isoform is not conserved outside of Drosophila. Overexpression experiments indicated that Loqs/R3D1-PA participates in endo-siRNA, but not exo-siRNA based silencing. We conclude that the functional specialization of Loqs-PD in Drosophila is a recently derived trait, and that in other dipterans, including the medically important mosquitoes, Loqs/R3D1-A participates in both the miRNA and endo-siRNA based pathways. 12 samples, 3 replicates of each type of sample. The GFP sample is the reference sample

ORGANISM(S): Aedes aegypti

SUBMITTER: Zach Adelman

PROVIDER: E-GEOD-65070 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

The hub protein loquacious connects the microRNA and short interfering RNA pathways in mosquitoes.

Haac Mary Etna ME Anderson Michelle A E MA Eggleston Heather H Myles Kevin M KM Adelman Zach N ZN

Nucleic acids research 20150312 7

Aedes aegypti mosquitoes vector several arboviruses of global health significance, including dengue viruses and chikungunya virus. RNA interference (RNAi) plays an important role in antiviral immunity, gene regulation and protection from transposable elements. Double-stranded RNA binding proteins (dsRBPs) are important for efficient RNAi; in Drosophila functional specialization of the miRNA, endo-siRNA and exo-siRNA pathway is aided by the dsRBPs Loquacious (Loqs-PB, Loqs-PD) and R2D2, respectiv ...[more]

PMID: 25765650

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Project description:Transposable elements are a serious threat for genome integrity and their control via small RNA mediated silencing pathways is an ancient strategy. The fruit fly Drosophila melanogaster has two silencing mechanisms that repress TEs expression: endogenous siRNAs (esiRNAs or endo-siRNAs) and Piwi-interacting small RNAs (piRNAs). The biogenesis of endo-siRNAs involves Loqs-PD, which acts predominantly during processing of dsRNA by Dcr-2, and R2D2 that primarily helps to direct siRNAs for loading into Ago2. We provide deep sequencing evidence consistent with the idea that R2D2 and Loqs-PD can function in part redundantly. Certain transposons display a preference for either dsRBD-protein for production or loading; this appeared to correlate neither with overall abundance, classification of the transposon or a specific site of genomic origin. The endo-siRNA biogenesis pathway in the germline operates according to the same principles as the existing model for the soma, and its impairment does not significantly affect piRNAs. Expanding the analysis, we confirmed the occurrence of somatic piRNA-like RNAs (pilRNAs) that show a ping-pong signature. We detected expression of the Piwi-family protein mRNAs only barely above background, indicating that the somatic pilRNAs may arise from a small sub-population of somatic cells that express a functional piRNA pathway. small RNA sampling experiment; small RNAs were prepared from head & thorax as well as dissected ovaries of Adult female Drosophila melanogaster. We used homozygous mutants of the dsRBD proteins Loqs and r2d2 to determine their contribution to the biogenesis of transposon-derived small RNAs. Heterozygous mutant animals served as control. For each RNA sample, we performed one deep-sequencing run without any treatment, and in parallel one sequencing run after periodate oxidation and beta-elimination. After this treatment, only Ago2, Piwi, Aub and Ago3-loaded small RNAs remain as they carry a 2'-O-methyl modification at their 3'-end. This helps to determine the loading status of the small RNAs detected. In total 8 different RNA samples were prepared and 16 libraries were sequenced.

Dataset Information

The hub protein Loqs connects the miRNA and endo-siRNA pathways in mosquitos

Publications

The hub protein loquacious connects the microRNA and short interfering RNA pathways in mosquitoes.

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