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• W3092098790 endingPage "e1008985" @default.
• W3092098790 startingPage "e1008985" @default.
• W3092098790 abstract "The arthropod melanization immune response is activated by extracellular protease cascades predominantly comprised of CLIP-domain serine proteases (CLIP-SPs) and serine protease homologs (CLIP-SPHs). In the malaria vector, Anopheles gambiae, the CLIP-SPHs SPCLIP1, CLIPA8, and CLIPA28 form the core of a hierarchical cascade downstream of mosquito complement that is required for microbial melanization. However, our understanding of the regulatory relationship of the CLIP-SPH cascade with the catalytic CLIP-SPs driving melanization is incomplete. Here, we report on the development of a novel screen to identify melanization pathway components based on the quantitation of melanotic mosquito excreta, eliminating the need for microdissections or hemolymph enzymatic assays. Using this screen, we identified CLIPC9 and subsequent functional analyses established that this protease is essential for the melanization of both Escherichia coli and the rodent malaria parasite Plasmodium berghei. Mechanistically, septic infection with E. coli promotes CLIPC9 cleavage and both full-length and cleaved CLIPC9 localize to this bacterium in a CLIPA8-dependent manner. The steady state level of CLIPC9 in the hemolymph is regulated by thioester-containing protein 1 (TEP1), suggesting it functions downstream of mosquito complement. In support, CLIPC9 cleavage is inhibited following SPCLIP1, CLIPA8, and CLIPA28 knockdown positioning it downstream of the CLIP-SPH cascade. Moreover, like CLIPA8 and CLIPA28, CLIPC9 processing is negatively regulated by serine protease inhibitor 2 (SRPN2). This report demonstrates how our novel excretion-based approach can be utilized to dissect the complex protease networks regulating mosquito melanization. Collectively, our findings establish that CLIPC9 is required for microbial melanization in An. gambiae and shed light on how the CLIP-SPH cascade regulates this potent immune response." @default.
• W3092098790 created "2020-10-15" @default.
• W3092098790 creator A5004227821 @default.
• W3092098790 creator A5009413614 @default.
• W3092098790 creator A5049467436 @default.
• W3092098790 creator A5069063077 @default.
• W3092098790 date "2020-10-12" @default.
• W3092098790 modified "2023-10-17" @default.
• W3092098790 title "The CLIP-domain serine protease CLIPC9 regulates melanization downstream of SPCLIP1, CLIPA8, and CLIPA28 in the malaria vector Anopheles gambiae" @default.
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• W3092098790 doi "https://doi.org/10.1371/journal.ppat.1008985" @default.
• W3092098790 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/7580898" @default.
• W3092098790 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/33045027" @default.
• W3092098790 hasPublicationYear "2020" @default.
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