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Integrated venom-gland transcriptomic and venom proteomic analysis of the spider Vacrothele uncata reveals the toxin diversity

ABSTRACT: This project conducts the first systematic analysis of the venom molecular diversity in the spider Vacrothele uncata. By integrating venom gland transcriptomics and venom proteomics, the study identifies 97 putative toxin transcripts and 84 expressed toxin proteins, including diverse disulfide-rich peptide neurotoxins (DRPs). This research elucidates the venom's molecular composition, supporting further understanding of the species' biological characteristics and laying the groundwork for discovering novel drug leads and bioinsecticides.

ORGANISM(S): Macrothele

SUBMITTER: Zizhong Yang

PROVIDER: PXD079221 | iProX | Wed Jun 03 00:00:00 BST 2026

REPOSITORIES: iProX

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Project description:Background The generalist dipteran pupal parasitoid Nasonia vitripennis injects 79 venom peptides into the host before egg laying. This venom induces several important changes in the host, including developmental arrest, immunosuppression, and alterations to normal metabolism. It is hoped that diverse and potent bioactivities of N. vitripennis venom provide an opportunity for the design of novel acting drugs. However, currently very little is known about the individual functions of N. vitripennis venom peptides and less than half can be bioinformatically annotated. The paucity of annotation information complicates the design of studies that seek to better understand the potential mechanisms underlying the envenomation response. Although the RNA interference system of N. vitripennis provides an opportunity to functionally characterise venom encoding genes, with 79 candidates this represents a daunting task. For this reason we were interested in determining the expression levels of venom encoding genes in the venom gland, such that this information could be used to rank candidate venoms. To do this we carried out deep sequencing of the transcriptome of the venom gland and neighbouring ovary tissue and used RNA-seq to measure expression from the 79 venom encoding genes. The generation of a specific venom gland transcriptome dataset also provides further opportunities to investigate novel features of this highly specialised organ. Results High throughput sequencing and RNA-seq revealed that the highest expressed venom encoding gene in the venom gland was a serine protease called Nasvi2EG007167, which has previously been implicated in the apoptotic activity of N. vitripennis venom. As expected the RNA-seq confirmed that the N. vitripennis venom encoding genes are almost exclusively expressed in the venom gland relative to the neighbouring ovary tissue. Novel peptides appear to perform key roles in N. vitripennis venom function as only four of the highest 15 expressed venom encoding genes are bioinformatically annotationed. The high throughput sequencing data also provided evidence for the existence of an additional 471 novel genes in the Nasonia genome that are expressed in the venom gland and ovary. Finally, metagenomic analysis of venom gland transcripts identified viral transcripts that may play an important part in the N. vitripennis venom function. Conclusions The expression level information provided here for the 79 venom encoding genes provides an unbiased dataset that can be used by the N. vitripennis community to identify high value candidates for further functional characterisation. These candidates represent bioactive peptides that have value in drug development pipelines.

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Integrated venom-gland transcriptomic and venom proteomic analysis of the spider Vacrothele uncata reveals the toxin diversity

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