Role of Clathrin Assembly Protein-2 Beta Subunit during White Spot Syndrome Virus Infection in Black Tiger Shrimp Penaeus monodon
Por:
Jatuyosporn, Thapanan, Laohawutthichai, Pasunee, Supungul, Premruethai, Sotelo-Mundo, Rogerio R., Ochoa-Leyva, Adrian, Tassanakajon, Anchalee, Krusong, Kuakarun
Publicada:
1 ene 2019
Categoría:
Multidisciplinary
Resumen:
White spot syndrome virus (WSSV) is one of the most lethal viruses severely affecting shrimp industry. This disease can cause 100% mortality of farmed shrimp within a week. This work aims to characterize clathrin assembly proteins in Penaeus monodon and investigate their roles in WSSV entry. In general, clathrin assembly proteins form complexes with specific receptors and clathrins, leading to clathrin-mediated endocytosis. Adaptor protein 2 (AP-2), which is responsible for endocytosis at plasma membrane, consists of four subunits including a, ß2, µ2 and s2. Knockdown of clathrin coat AP17, or s subunit of AP-2 dramatically reduced WSSV infectivity. Similar results were observed, when shrimp were pre-treated with chlorpromazine (CPZ), an inhibitor of clathrin-dependent endocytosis. The complete open reading frames of AP-2ß and µ subunits of P. monodon are reported. PmAP-2 ß was up-regulated about 4-fold at 6 and 36 h post-WSSV infection. Knockdown of PmAP-2ß delayed shrimp mortality during WSSV infection, of which WSSV intermediate early 1 gene expression was also down-regulated. Immunogold-labelling and transmission electron microscopy revealed that PmAP-2ß co-localized with WSSV particles at plasma membrane. In addition, PmAP-2ß-silencing significantly affected the expression levels of PmSTAT, PmDOME, PmDorsal and ALFPm3 during WSSV infection. It is possible that PmAP-2ß is associated with the JAK/STAT and the Toll pathway.
Filiaciones:
Structural and Computational Biology Research Unit, Department of Biochemistry, Faculty of Science, Chulalongkorn UniversityBangkok 10330, Thailand
Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn UniversityBangkok 10330, Thailand
National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathumthani12120, Thailand
Laboratorio de Estructura Biomolecular, Centro de Investigación en Alimentación Y Desarrollo, A.C. (CIAD), Carretera Gustavo Enrique Astiazaran Rosas No. 46, Hermosillo, Sonora 83304, Mexico
Departamentos de Microbiología Molecular, Medicina Molecular y Bioprocesos, Unidad Universitaria de Secuenciacián Masiva y Bioinformática, Instituto de Biotecnología (IBT), Universidad Nacional Autónoma de México (UNAM), Avenida Universidad 2001 ,Colonia Chamilpa, Cuernavaca, 62210, Mexico
Chulalongkorn Univ, Fac Sci, Dept Biochem, Struct & Computat Biol Res Unit, Bangkok 10330, Thailand
Chulalongkorn Univ, Fac Sci, Dept Biochem, Ctr Excellence Mol Biol & Genom Shrimp, Bangkok 10330, Thailand
NSTDA, Natl Ctr Genet Engn & Biotechnol BIOTEC, Pathum Thani 12120, Thailand
Ctr Invest Alimentac & Desarrollo AC CIAD, Lab Estruct Biomol, Carretera Gustavo Enrique Astiazaran Rosas 46, Hermosillo 83304, Sonora, Mexico
UNAM, Inst Biotecnol IBT, Unidad Univ Secuenciacian Mas & Bioinformat, Dept Microbiol Mol, Ave Univ 2001, Cuernavaca 62210, Morelos, Mexico
UNAM, Inst Biotecnol IBT, Unidad Univ Secuenciacian Mas & Bioinformat, Dept Med Mol, Ave Univ 2001, Cuernavaca 62210, Morelos, Mexico
UNAM, Inst Biotecnol IBT, Unidad Univ Secuenciacian Mas & Bioinformat, Dept Bioproc, Ave Univ 2001, Cuernavaca 62210, Morelos, Mexico
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