Project description:We report the cloning and sequencing of both endogenous small RNAs and virus-derived siRNAs produced by the antiviral RNAi pathway in Drosophila. We find that a diverse panel of viruses are targeted by the RNAi pathway in Drosophila to produce abundant virus-derived siRNAs, and these siRNAs map to various locations within the viral genomes. Knockdown of various RNAi and miRNA pathway components alters the levels of these viral small RNAs.
Project description:High throughput seqeuncing of small RNAs (PAGE isolated from total RNA or Argonaute immunoprecipitates) from Drosophila melanogaster using the Illumina platform. Adapter ligation requires 5' monophosphate and 3' OH. Full analysis of all libraries in this set is published (Czech B. et al. 2008), leading to the description of endogenous siRNAs in flies. Keywords: Solexa sequences
Project description:Proteomic Analysis (MS/MS) of Drosophila melanogaster mtx2 (Ortholog of CG8004) Heterozygous versus Homozygous Mutants at 2 Days Post-Pupa Formation
Project description:<p>Viral studies of Drosophila melanogaster typically involve virus injection with a small needle, causing post-injury a wounding/wound healing response, in addition to the effects of viral infection. However, the metabolic response to the needle injury is understudied, and many viral investigations neglect potential effects of this response. Furthermore, the wMel strain of the endosymbiont bacterium Wolbachia pipientis provides anti-viral protection in Drosophila. Here we used NMR-based metabolomics to characterise the acute wounding response in Drosophila and the relationship between wound healing and the Wolbachia strain wMel. The most notable response to wounding was found on the initial day of injury and lessened with time in both uninfected and Wolbachia infected flies. Metabolic changes in injured flies revealed evidence of inflammation, Warburg-like metabolism and the melanisation immune response as a response to wounding. In addition, at five days post injury Wolbachia infected injured flies were metabolically more similar to the uninjured flies than uninfected injured flies were at the same time point, indicating a positive interaction between Wolbachia infection and wound healing. This study is the first metabolomic characterisation of the wound response in Drosophila and its findings are crucial to the metabolic interpretation of viral experiments in Drosophila in both past and future studies.</p>
Project description:Cis-natural antisense transcripts (cis-NATs) have been speculated to be substrates for endogenous RNA interference (RNAi), but little experimental evidence for such a pathway in animals has been reported. Analysis of massive Drosophila melanogaster small RNA data sets now reveals that endogenous small interfering RNAs (siRNAs) are produced via bidirectional transcription. >100 cis-NATs with overlapping 3' exons generate 21-nt, Dicer-2 (Dcr-2)dependent, 3'-end modified siRNAs. To determine whether any co-expressed cisNATs are denied entry into the RNAi pathway, we analyzed the gene expression profile of S2 cells. The analysis suggested that the processing of cis-NATs by RNAi are actively restricted, and the selected loci are enriched for nucleic acidbased functions and include Argonaute-2 (AGO2) itself. Keywords: Gene expression
Project description:Thermal acclimation study on Drosophila melanogaster reared at 3 different temperatures (12, 25, and 31oC). The proteomic profiles of D. melanogaster under these different temperatures were analyzed and compared using label-free tandem mass spectrometry.
