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Proc. Natl. Acad. Sci. USA, Jun 25 2024, 121 (26) | https://doi.org/10.1073/pnas.2322927121
StaufenC facilitates utilization of the ERAD pathway to transport dsRNA through the endoplasmic reticulum to the cytosol
Jinmo Kooa,1 and Subba Reddy Pallia,2
aDepartment of Entomology, Collegeof Agriculture, University of Kentucky, Lexington, KY 40546
1Present address: Department of Entomology, Universityof California, Riverside, CA 92521.
2To whom correspondence may be addressed.: Subba Reddy Palli
Abstract
RNA interference (RNAi) is more efficient in coleopteran insects than other insects. StaufenC (StauC), a coleopteran-specific double-stranded RNA (dsRNA)-binding protein, is required for efficient RNAi in coleopterans. We investigated the function of StauC in the intracellular transport of dsRNA into the cytosol, where dsRNA is digested by Dicer enzymes and recruited by Argonauts to RNA-induced silencing complexes. Confocal microscopy and cellular organelle fractionation studies have shown that dsRNA is trafficked through the endoplasmic reticulum (ER) in coleopteran Colorado potato beetle (CPB) cells. StauC is localized to the ER in CPB cells, and StauC-knockdown caused the accumulation of dsRNA in the ER and a decrease in the cytosol, suggesting that StauC plays a key role in the intracellular transport of dsRNA through the ER. Using immunoprecipitation, we showed that StauC is required for dsRNA interaction with ER proteins in the ER-associated protein degradation (ERAD) pathway, and these interactions are required for RNAi in CPB cells. These results suggest that StauC works with the ERAD pathway to transport dsRNA through the ER to the cytosol. This information could be used to develop dsRNA delivery methods aimed at improving RNAi.
논문정보
- Research article
- 2024년 06월 (BRIC 등록일 2024-06-19)
- 국외 연구진
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