Rab7 GTPase, a direct target of miR-131-3p, limits intracellular Spiroplasma eriocheiris infection by modulating phagocytosis

Spiroplasma eriocheiris is a kind of intracellular pathogen without cell wall and the causative agent of Chinese mitten crab Eriocheir sinensis “tremor disease”, which causes significant economic losses in the crustacean aquaculture. However, little is known about the intracellular transport of this pathogen and host innate immune response to this pathogen. Rab GTPases are key regulators for endocytosis and intracellular pathogen trafficking. In this study, we showed that S. eriocheiris infection upregulated the transcription of Rab7 through the downregulation of miR-131-3p. Subsequently, both hemocytes transfected with miR-131-3p mimics and hemocytes derived from Rab7 knockdown crabs exhibited reduced phagocytic activities and increased susceptibility to S. eriocheiris infection. Additionally, Rab7 could interact with the cell shape-determining protein MreB3 of S. eriocheiris, and its overexpression promoted S. eriocheiris internalization and fusion with lysosomes, thereby limiting S. eriocheiris replication in Drosophila S2 cells. Overall, these results demonstrated that Rab7 facilitated host cell phagocytosis and interacted with MreB3 of S. eriocheiris to prevent S. eriocheiris infection. Moreover, miR-131-3p was identified as a negative regulator of this process through its targeting of Rab7. Therefore, targeting miR-131-3p might be an effective strategy for controlling S. eriocheiris in crab aquaculture. •E. sinensis Rab7 and miR-131-3p were changed after S. eriocheiris infection.•EsRab7 was target gene of miR-131-3p.•The knockdown of EsRab7 reduced phagocytosis and increased S. eriocheiris infection.•EsRab7 could interact with S. eriocheiris MreB3 to promote S. eriocheiris internalization.•Overexpression of EsRab7 limited S. eriocheiris replication in Drosophila S2 cells.