Dataset Information

Characterization of a cell death-inducing endonuclease-like venom protein from the parasitoid wasp Pteromalus puparum (Hymenoptera: Pteromalidae).

ABSTRACT:

Background

Parasitoid wasps are valuable natural enemies for controlling pests. To ensure successful parasitism, these wasps inject venoms along with their eggs that are deposited either into or on their hosts. Parasitoid venoms regulate host behaviors, development, metabolism and immune responses. Pteromalus puparum is a pupal endoparasitoid that parasitizes a number of butterflies, including the worldwide pest cabbage butterfly, Pieris rapae. Venom from P. puparum has a variety of effects on host hemocytes, including alteration of absolute and relative hemocyte counts, and inhibition of hemocyte spreading and encapsulation. In particular, P. puparum venom causes hemocyte cell death in vivo and in vitro.

Results

Using assay-guided chromatography, a cell death-inducing venom fraction was identified and defined as P. puparum endonuclease-like venom protein (PpENVP). It belongs to the DNA/RNA nonspecific endonuclease family, which contains two conserved endonuclease activation sites. We analyzed its expression profiles and demonstrated that PpENVP inhibits gene expression in transfected cells relying on two activation sites. However, RNA interference of PpENVP did not significantly reduce P. puparum venom cytotoxicity, suggesting that PpENVP may not be the sole cytotoxic factor present.

Conclusion

Our results provide novel insight into the function of the P. puparum venom cocktail and identify a promising insecticide candidate endonuclease that targets insect hemocytes.

SUBMITTER: Wang J

PROVIDER: S-EPMC9282878 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundInsect hosts have evolved immunity against invasion by parasitoids, and in co-evolutionary response parasitoids have also developed strategies to overcome host immune systems. The mechanisms through which parasitoid venoms disrupt the promotion of host immunity are still unclear. We report here a new mechanism evolved by parasitoid Pteromalus puparum, whose venom inhibited the promotion of immunity in its host Pieris rapae (cabbage white butterfly).Methodology/principal findingsA full-length cDNA encoding a C-type lectin (Pr-CTL) was isolated from P. rapae. Quantitative PCR and immunoblotting showed that injection of bacterial and inert beads induced expression of Pr-CTL, with peaks of mRNA and Pr-CTL protein levels at 4 and 8 h post beads challenge, respectively. In contrast, parasitoid venom suppressed Pr-CTL expression when co-injected with beads, in a time and dose-dependent manner. Immunolocalization and immunoblotting results showed that Pr-CTL was first detectable in vesicles present in cytoplasm of granulocytes in host hemolymph, and was then secreted from cells into circulatory fluid. Finally, the secreted Pr-CTL bound to cellular membranes of both granulocytes and plasmatocytes. Injection of double-stranded RNA specific for target gene decreased expression of Pr-CTL, and a few other host immune-related genes. Suppression of Pr-CTL expression also down-regulated antimicrobial and phenoloxidase activities, and reducing phagocytotic and encapsulation rates in host. The inhibitory effect of parasitoid venom on host encapsulation is consistent with its effect in suppressing Pr-CTL expression. Binding assay results showed that recombinant Pr-CTL directly attached to the surface of P. puparum egges. We infer that Pr-CTL may serve as an immune signalling co-effector, first binding to parasitoid eggs, regulating expression of a set of immune-related genes and promoting host immunity.Conclusions/significanceP. puparum venom inhibits promotion of host immune responses by silencing expression of host C-type lectin gene Pr-CTL, whose expression affected transcription of other host immune-related genes.