Project description:While the unique symbiotic relationship between anemonefish and sea anemones is iconic, it is still not fully understood how anemonefish withstand and thrive within this venomous host environment. In this study we used a proteotranscriptomics approach to elucidate the proteinaceous toxin repertoire from the most popular host sea anemone Entacmaea quadricolor. Although 1251 different toxin or toxin-like RNA transcripts were expressed in E.quadricolor tentacles and 2736 proteins were detected in milked venom, only 135 (approx. 10%) of proteins in venom were classified as putative toxins. This work raises the perils of defining a dominant venom type based on transcriptomics data alone in sea anemones, as we found that the dominant venom type differed between the transcriptome and proteome data. Moreover, anemonefishes interact with sea anemone proteins, so it is important when determining the dominant toxin type to examine the peptides and proteins that are present in host sea anemone venom and mucus which anemonefishes are known to interact.
Project description:Both single cell and bulk RNA sequencing was performed on expanding or differentiating snake venom gland organoids (from Aspidelaps Lubricus Cowlesi and Naja Nivea), or tissue (Aspidelaps Lubricus Cowlesi). Bulk RNA sequencing from the snake venom gland, liver and pancreas was performed to construct a de novo transcriptome using Trinity.
Project description:Diachasmimorpha longicaudata parasitoid wasps carry a symbiotic poxvirus, known as DlEPV, within the female wasp venom gland. We sequenced RNA from venom gland tissue to identify DlEPV orthologs for 3 conserved poxvirus core genes. The DlEPV ORFs identified from this transcriptome were used to design primers for downstream RT-qPCR analysis and RNAi knockdown experiments.
Project description:Venoms from marine animals have been recognized as a new emerging source of peptide-based therapeutics. Several peptide toxins from sea anemone have been investigated as therapeutic leads or pharmacological tools. Venom complexity should be further confirmed using novel strategies of large-scale sequencing which integrated transcriptomics and proteomics to provide new putative proteins or peptides to be compared among species. We applied transcriptomic combining with proteomic analyses and identified six groups of peptide toxins including neurotoxin, hemostatic and hemorrhagic toxin, protease inhibitor, mixed function enzymes, venom auxiliary proteins, and peptides related to allergen and innate immunity.
Project description:Entomopathogenic nematodes (EPNs) are unique parasitic nematodes due to their symbiosis with entomopathogenic bacteria and their ability to kill insect hosts quickly after infection. Although it has been widely believed that EPNs rely on their bacterial partners for killing insect hosts, compelling evidence from previous studies challenges this model. We developed an improved method of activating millions of Steinernema carpocapsae infective juveniles (IJs) in vitro to harvest excreted/secreted (ES) proteins for bioactivity tests and proteomics analysis. We found that a low dose of the ES proteins from early activated nematodes is lethal to Drosophila melanogaster adults within 2-6 hours. We analyzed the protein composition of this venom using mass spectrometry and identified 472 proteins. Many of these venom proteins share high homology with those of vertebrate-parasitic nematodes. Among many different families of proteins found in the venom, proteases and protease inhibitors are especially abundant. Some toxin-related proteins such as Shk domain-containing proteins were also detected. We further analyzed the transcriptomes of individual non-activated IJs and nematodes that were activated in vitro and in vivo, which revealed a dramatic shift in gene expression during IJ activation. By comparing the whole transcriptomes and the genes encoding venom proteins between the in vitro and in vivo activated nematodes, we confirmed that the in vitro activation is a good approximation of the in vivo process. In summary, our findings strongly support a new model that S. carpocapsae and likely other Steinernema EPNs have a more active role in contributing to the pathogenicity of the nematode-bacterium complex than simply relying on their symbiotic bacteria. Furthermore, we propose that EPNs are a good model system for investigating vertebrate- and human-parasitic nematodes, especially regarding the function of ES products.