title: White spot syndrome virus entry is dependent on multiple endocytic routes and strongly facilitated by Cq-GABARAP in a CME-dependent manner.
abstract: White spot syndrome virus (WSSV) is a lethal pathogen of shrimp and many other crustaceans, including crayfish. However, the molecular mechanism underlying its cellular entry remains elusive due to the lack of shrimp cell lines for viral propagation. Crayfish hematopoietic tissue (Hpt) cell culture was recently established as a good model for WSSV infection study. Here, we showed that multiple endocytic routes, including clathrin-mediated endocytosis (CME), macropinocytosis and caveolae-mediated endocytosis, were indispensably employed for the viral entry into Hpt cell of the crayfish Cherax quadricarinatus. Intriguingly, cellular autophagic activity was positively correlated with efficient viral entry, in which a key autophagy-related protein, γ-aminobutyric acid receptor-associated protein (Cq-GABARAP), that not only localized but also co-localized with WSSV on the Hpt cell membrane, strongly facilitated WSSV entry by binding to the viral envelope VP28 in a CME-dependent manner that was negatively regulated by Cq-Rac1. Furthermore, cytoskeletal components, including Cq-β-tubulin and Cq-β-actin, bound to both recombinant rCq-GABARAP and WSSV envelope proteins, which likely led to viral entry promotion via cooperation with rCq-GABARAP. Even under conditions that promoted viral entry, rCq-GABARAP significantly reduced viral replication at an early stage of infection, which was probably caused by the formation of WSSV aggregates in the cytoplasm.
authors.affiliation: State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361102, Fujian, PR China.
authors.forename: Rong-Yuan
authors.initials: RY
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361102, Fujian, PR China.
authors.forename: Kai-Li
authors.initials: KL
authors.lastname: Shen
authors.affiliation: State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361102, Fujian, PR China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361102, Fujian, PR China.|Faculty of Life Science and Biotechnology, Ningbo University, 818 Fenghua Road, Ningbo 315211, Zhejiang, PR China.
authors.forename: Wei-Wei
authors.initials: WW
authors.lastname: Fan
authors.affiliation: State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361102, Fujian, PR China.
authors.forename: Xiao-Lu
authors.initials: XL
authors.lastname: Xie
authors.affiliation: State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361102, Fujian, PR China.
authors.forename: Chuang
authors.initials: C
authors.lastname: Meng
authors.affiliation: State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361102, Fujian, PR China.
authors.forename: Xue-Jiao
authors.initials: XJ
authors.lastname: Chang
authors.affiliation: State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361102, Fujian, PR China.
authors.forename: Li-Bing
authors.initials: LB
authors.lastname: Zheng
authors.affiliation: State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361102, Fujian, PR China.
authors.forename: Joseph
authors.initials: J
authors.lastname: Jeswin
authors.affiliation: Faculty of Life Science and Biotechnology, Ningbo University, 818 Fenghua Road, Ningbo 315211, Zhejiang, PR China.
authors.forename: Cheng-Hua
authors.initials: CH
authors.lastname: Li
authors.affiliation: State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361102, Fujian, PR China.|Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources, State-Province Joint Engineering Laboratory of Marine Bioproducts and Technology, Xiamen 361102, Fujian, PR China.
authors.forename: Ke-Jian
authors.initials: KJ
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361102, Fujian, PR China.|Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources, State-Province Joint Engineering Laboratory of Marine Bioproducts and Technology, Xiamen 361102, Fujian, PR China.
authors.forename: Hai-Peng
authors.initials: HP
authors.lastname: Liu
mesh_terms: D000818:Animals; D060829:Arthropod Proteins / Q000502:physiology*; D003400:Astacoidea / Q000166:cytology / Q000821:virology; D001343:Autophagy; D000071190:Autophagy-Related Protein 8 Family / Q000502:physiology*; D002478:Cells, Cultured; D003034:Coated Pits, Cell-Membrane / Q000648:ultrastructure / Q000821:virology; D004705:Endocytosis; D011485:Protein Binding; D014759:Viral Envelope Proteins / Q000378:metabolism; D053586:Virus Internalization; D014779:Virus Replication; D046848:White spot syndrome virus 1 / Q000502:physiology*
chemical_list: D060829:Arthropod Proteins; D000071190:Autophagy-Related Protein 8 Family; D014759:Viral Envelope Proteins