title: White spot syndrome virus enters crayfish hematopoietic tissue cells via clathrin-mediated endocytosis.
abstract: White spot syndrome virus (WSSV) is a major pathogen of aquacultured shrimp. However, the mechanism of its entry remains poorly understood. In this study, by analyzing the internalization of WSSV using crayfish hematopoietic tissue (HPT) cells, we showed that WSSV virions were engulfed by cell membrane invaginations sharing the features of clathrin-coated pits and then internalized into coated cytoplasmic vesicles. Further investigation indicated that WSSV internalization was significantly inhibited by chlorpromazine (CPZ) but not genistein. The internalized virions were colocalized with endogenous clathrin as well as transferrin which undergoes clathrin-dependent uptake. Preventing endosome acidification by ammonium chloride (NH4Cl) or chloroquine (CQ) dramatically reduced WSSV entry as well. Moreover, disturbance of dynamin activity or depletion of membrane cholesterol also blocked WSSV uptake. These data indicate that WSSV enters crayfish HPT cells via clathrin-mediated endocytosis in a pH-dependent manner, and membrane cholesterol as well as dynamin is critical for efficient viral entry.
authors.affiliation: Key Laboratory of Marine Genetic Resources, South China Sea Bio-Resource Exploitation and Utilization Collaborative Innovation Center, Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, China.
authors.forename: Jiajun
authors.initials: J
authors.lastname: Huang
authors.affiliation: Key Laboratory of Marine Genetic Resources, South China Sea Bio-Resource Exploitation and Utilization Collaborative Innovation Center, Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, China. Electronic address: lifang_87@163.com.
authors.forename: Fang
authors.initials: F
authors.lastname: Li
authors.affiliation: Key Laboratory of Marine Genetic Resources, South China Sea Bio-Resource Exploitation and Utilization Collaborative Innovation Center, Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, China.
authors.forename: Junjun
authors.initials: J
authors.lastname: Wu
authors.affiliation: Key Laboratory of Marine Genetic Resources, South China Sea Bio-Resource Exploitation and Utilization Collaborative Innovation Center, Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, China. Electronic address: yangfeng5274@163.com.
authors.forename: Feng
authors.initials: F
authors.lastname: Yang
mesh_terms: D000818:Animals; D003400:Astacoidea / Q000502:physiology / Q000821:virology*; D002784:Cholesterol / Q000378:metabolism; D002966:Clathrin / Q000378:metabolism; D004705:Endocytosis; D006413:Hematopoietic System / Q000821:virology; D053586:Virus Internalization; D046848:White spot syndrome virus 1 / Q000502:physiology*
chemical_list: D002966:Clathrin; D002784:Cholesterol
keywords: Clathrin-mediated endocytosis; Entry; White spot syndrome virus