The dual roles of Armigeres subalbatus prophenoloxidase V in parasite melanization and egg chorion melanization in the mosquito Ar. subalbatus |
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| Affiliation: | 1. Institute of Microbiology and Immunology, National Yang-Ming University, Shih-Pai, Taipei 112, Taiwan;2. Department of Pathobiological Sciences, 1656 Linden Drive, University of Wisconsin, Madison, WI 53706, USA;1. Eszterházy Károly College, Institute of Food Science, Leányka u. 6., Eger H-3300, Hungary;2. Central Environmental and Food Science Research Institute, Herman Ottó u. 15., Budapest H-1022, Hungary;1. Nomis Center for Immunobiology and Microbial Pathogenesis, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA;1. Department of Biomedicine, Aarhus University, DK-8000 Aarhus C, Denmark;2. Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Jagtvej 162,, DK-2100 Copenhagen, Denmark;3. Department of Molecular Biology and Genetics, Aarhus University, DK-8000 Aarhus C, Denmark;4. NeuroSearch A/S, Pederstrupvej 93, DK-2750 Ballerup, Denmark;5. H. Lundbeck A/S, Ottiliavej 9, DK-2500 Valby, Denmark;1. Section on Physical Biochemistry, Laboratory of Biochemistry and Genetics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland |
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| Abstract: | Phenoloxidases (POs) play key roles in various physiological functions in insects, e.g., cuticular sclerotization, wound healing, egg tanning, cuticle formation and melanotic encapsulaction of pathogens. Previously, we identified five POs, designated As-pro-PO I–V, from the mosquito Armigeres subalbatus and demonstrated that the functions of As-pro-PO I, II and III, were associated with filarial parasite melanization, blood feeding and cuticle formation, respectively. In the present study, we delineate the dual functions of As-pro-PO V. We found that the level of As-pro-PO V mRNA in mosquitoes was significantly increased after microfilaria challenge or blood feeding, and decreased to normal level after oviposition. Knockdown of As-pro-PO V by dsRNA resulted in significant decreases in the degree of microfilaria melanization, egg chronic melanization rates and egg hatching rates in Ar. subalbatus. Further transfection and electrophoretic mobility-shift assays verified the As-pro-PO V gene might regulated by both AP-1, a putative immune-related regulatory element and CdxA, a developmental regulatory element. The binding of AP-1 and CdxA motif with mosquito nuclear extracts was significantly enhanced after microfilaria challenge and blood-feeding in Ar. subalbatus, respectively. These results indicate that As-pro-PO V is a critical enzyme that is required for both an effective melanization immune response and egg chorion melanization in this mosquito. |
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| Keywords: | Mosquito Prophenoloxidase Double stranded RNA-mediated RNAi Parasite melanization Egg chorion melanization Gene regulation |
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