An ATP synthase beta subunit is required for internalization of dsRNA into shrimp cells

Extracellular double-stranded RNA (dsRNA) is an important modulator in innate immunity in both vertebrates and invertebrates. In shrimp, extracellular dsRNA can trigger RNAi pathway and serves as antiviral defense mechanism. However, the mechanism of dsRNA internalization into the cells has not yet known in shrimp cells. This study identified candidate cell surface proteins from shrimp hepatopancreatic cells that could interact with dsRNA by a ligand blot assay. Among the candidate proteins, a cell-surface beta subunit of ATP synthase was shown to be capable of internalizing dsRNA into shrimp hepatopancreatic cells that could rapidly occur in just 1 min upon dsRNA challenge. Colocalization between dsRNA and ATP synthase beta subunit implied correlation between dsRNA and ATP synthase beta subunit during dsRNA internalization. Furthermore, dsRNA showed colocalization with Ras-related endocytic proteins, Rab5 and Rab7 indicating that dsRNA was internalized via the receptor-mediated endocytosis. For the above evidences as well as the reduction of dsRNA internalization by angiostatin and antibodies against ATP synthase beta subunit, we propose that dsRNA interacts with ATP synthase via a nucleotide binding site in the beta subunit prior to internalize dsRNA into cells.