Project description:We report the application of Cappable-seq to selectively enrich prokaryotic endosymbiont transcripts from mixed host-symbiont total RNA.
Project description:Brugia malayi is a parasitic nematode that causes lymphatic filariasis in humans. A total of 178 novel microRNA were identified from short read transcriptional data, which when combined with known Brugia microRNAs yielded a total of 284 microRNA. Of these, 123 microRNA sequences (43%) are differentially expressed over the mammalian life stages of B. malayi that we examined. Putative targets of these microRNA were identified from inversely expressed target clusters that contain valid seed sequences for the corresponding microRNAs. The largest identified cluster is downregulated in adult females and enriched in zinc finger domains, helicase domains, and DNA binding domains suggesting this microRNA cluster may have regulatory control over a large proportion of adult female specific mRNA genes. MicroRNA-like molecules are identified as produced by the Wolbachia endosymbiont, providing evidence for direct nucleic acid-based interdomain communication between filarial nematodes and their bacterial obligate endosymbiont.
Project description:Wolbachia pipientis is a ubiquitous intracellular bacterium that is known for its manipulation of reproduction in arthropod hosts. Wolbachia has also been shown to colonize virtually all somatic tissues, including the brain, but little is known about the interaction between host and bacterium in these locations. To this end, we studied the effects of Wolbachia infection on the brain of Drosophila melanogaster. Using comparative proteomics, we uncovered the post-translational modification of many proteins within the Drosophila head and body upon infection, with glutamic acid decarboxylase being modified within the head only. Given this enzyme’s role in neurotransmitter synthesis, we next tested how Wolbachia infection impacts various behaviors and GABA production within Drosophila. We discovered an improved response to yeast odors in Wolbachia-infected, mated females compared to their uninfected counterparts. Gross measurements of GABA in whole brains showed no detectable change in GABA abundance upon infection. Treatments with GABA agonist indicated that the behavioral change was not GABA-dependent, leaving the mechanism behind Wolbachia-mediated changes in behavior obscure. Given the multiple protein changes in the Drosophila head upon infection, we propose a model in which Wolbachia drives the modification of glutamic acid decarboxylase, and several metabolic proteins, to increase survival in the specialized niche of the brain. These results give rise to new questions about the Wolbachia-Drosophila relationship and future work will focus on the mechanism through which Wolbachia confers these protein changes.
Project description:We sequenced total RNA from Dirofilaria immitis in order to generate the first tissue-specific gene expression profile of a filarial nematode and its Wolbachia endosymbiont.