Early endocytosis pathways in SSN-1 cells infected by dragon grouper nervous necrosis virus.
Many fish undergo betanodavirus infection. To study the infection process of dragon grouper nervous necrosis virus (DGNNV), native virus and virus-like particles (VLPs) were used to analyse the binding and internalization in SSN-1 cells. The binding of DGNNV and VLPs to SSN-1 cells was demonstrated using Western blotting and immunofluorescence microscopy. As estimated by indirect ELISA, the DGNNV particles bound SSN-1 cells in a dose-dependent manner up to 8 x 10(4) particles per cell. The binding of VLPs was sensitive to neuraminidase and tunicamycin, suggesting that cell-surface sialic acid is involved in binding. The penetration of DGNNV into cells, which was monitored by electron microscopy, appeared to occur mainly via the spherical pit and membrane ruffling pathways. Occasionally, a spherical pit was engulfed by membrane ruffling so as to form a large figure-of-eight-shaped vesicle with an open connection. Our observations suggest that DGNNV utilizes both micro- and macropinocytosis pathways to enter SSN-1 cells.
Liu
Wangta
W
Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 804, Taiwan.
Hsu
Chi-Hsin
CH
Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 804, Taiwan.
Hong
Yi-Ren
YR
Graduate Institute of Biochemistry, Kaohsiung Medical University, Kaohsiung, Taiwan.
Wu
Shu-Chuan
SC
Graduate Institute of Biochemistry, Kaohsiung Medical University, Kaohsiung, Taiwan.
Wang
Chun-Hsiung
CH
Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 804, Taiwan.
Wu
Yi-Min
YM
Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 804, Taiwan.
Chao
Chia-Ben
CB
Institute for Animal Disease Prevention and Control, Kaohsiung, Taiwan.
Lin
Chan-Shing
CS
Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 804, Taiwan.
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D004705:Endocytosis / Q000502:physiology*; D005399:Fishes / Q000821:virology*; D016253:Microscopy, Immunoelectron; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D030381:Nodaviridae / Q000378:metabolism / Q000472:pathogenicity* / Q000648:ultrastructure; D014771:Virion / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D014779:Virus Replication
D019158:N-Acetylneuraminic Acid