C-terminal domain on the outer surface of the Macrobrachium rosenbergii nodavirus capsid is required for Sf9 cell binding and internalization.
We have shown that Macrobrachium rosenbergii nodavirus (MrNV) was able to infect Sf9 cells and that MrNV virus-like particles (MrNV-VLPs) were capable nanocontainers for delivering nucleic acid-based materials. Here, we demonstrated that chymotryptic removal of a C-terminal peptide and its truncated variant (F344-MrNV-VLPs) exhibited a drastically reduced ability to interact and internalize into Sf9 cells. Electron microscopic observations revealed that the loss of C-terminal domain either from enzyme hydrolysis or genetic truncation did not affect the generated MrNV-VLPs' icosahedral conformation, but did drastically affect the VLPs' internalization ability into Sf9 cells. Homology-based modelling of the MrNV capsid with other icosahedral capsid models revealed that this chymotrypsin-sensitive C-terminal domain was not only exposed on the capsid surface, but also constituted the core of the viral capsid protrusion. These results therefore suggest the importance of the C-terminal domain as a structure for targeted cell interaction which is presumably localized at the protruding domain. This work thus provided the functional insights into the role of the MrNV C-terminal domain in viral entry into Sf9 cells and lead to the development of strategies in combatting MrNV infection in susceptible cells.
Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Road, Ratchathewi, Bangkok, 10400, Thailand.
Monsicha
M
Somrit
Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Road, Ratchathewi, Bangkok, 10400, Thailand; Department of Anatomy, Faculty of Medicine, Srinakharinwirot University, Bangkok, 10110, Thailand.
Atthaboon
A
Watthammawut
Center of Excellence for Shrimp Molecular Biology and Biotechnology, Faculty of Science, Mahidol University, Rama VI Road, Ratchathewi, Bangkok, 10400, Thailand; Nakhonsawan campus, Mahidol University, Nakhonsawan 60130, Thailand.
Charoonroj
C
Chotwiwatthanakun
Department of Biology, Faculty of Science, Mahidol University, Rama VI Road, Ratchathewi, Bangkok, 10400, Thailand; Center of Excellence on Environmental Health and Toxicology, Office of Higher Education Commission, Ministry of Education, Bangkok, 10400, Thailand.
Puey
P
Ounjai
Center of Excellence for Shrimp Molecular Biology and Biotechnology, Faculty of Science, Mahidol University, Rama VI Road, Ratchathewi, Bangkok, 10400, Thailand.
Wanida
W
Suntimanawong
Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Road, Ratchathewi, Bangkok, 10400, Thailand. Electronic address: wattana.wee@mahidol.ac.th.
Wattana
W
Weerachatyanukul
D000595:Amino Acid Sequence; D000818:Animals; D002213:Capsid / Q000737:chemistry / Q000378:metabolism*; D005434:Flow Cytometry; D054884:Host-Pathogen Interactions; D008958:Models, Molecular; D030381:Nodaviridae / Q000502:physiology* / Q000648:ultrastructure; D033541:Palaemonidae / Q000821:virology*; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D061987:Sf9 Cells; D019065:Virus Assembly; D053585:Virus Attachment; D053586:Virus Internalization
Internalization; Nodavirus; Shrimp; VLP; Virus-host interaction