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Inclusion of an Arg-Gly-Asp receptor-recognition motif into the capsid protein of rabbit hemorrhagic disease virus enables culture of the virus
- Source :
- The Journal of biological chemistry. 292(21)
- Publication Year :
- 2017
-
Abstract
- The fact that rabbit hemorrhagic disease virus (RHDV), an important member of the Caliciviridae family, cannot be propagated in vitro has greatly impeded the progress of investigations into the mechanisms of pathogenesis, translation, and replication of this and related viruses. In this study, we have successfully bypassed this obstacle by constructing a mutant RHDV (mRHDV) by using a reverse genetics technique. By changing two amino acids (S305R,N307D), we produced a specific receptor-recognition motif (Arg-Gly-Asp; called RGD) on the surface of the RHDV capsid protein. mRHDV was recognized by the intrinsic membrane receptor (integrin) of the RK-13 cells, which then gained entry and proliferated as well as imparted apparent cytopathic effects. After 20 passages, the titers of RHDV reached 1 × 104.3 50% tissue culture infectious dose (TCID50)/ml at 72 h. Furthermore, mRHDV-infected rabbits showed typical rabbit plague symptoms and died within 48–72 h. After immunization with inactivated mRHDV, the rabbits survived wild-type RHDV infection, indicating that mRHDV could be a candidate virus strain for producing a vaccine against RHDV infection. In summary, this study offers a novel strategy for overcoming the challenges of proliferating RHDV in vitro. Because virus uptake via specific membrane receptors, several of which specifically bind to the RGD peptide motif, is a common feature of host cells, we believe that this the strategy could also be applied to other RNA viruses that currently lack suitable cell lines for propagation such as hepatitis E virus and norovirus.
- Subjects :
- 0301 basic medicine
Virus Cultivation
Receptors, Peptide
040301 veterinary sciences
Viral protein
Hemorrhagic Disease Virus, Rabbit
Amino Acid Motifs
Biology
medicine.disease_cause
Biochemistry
Microbiology
Virus
Madin Darby Canine Kidney Cells
0403 veterinary science
03 medical and health sciences
Dogs
Hepatitis E virus
Viral entry
Cricetinae
Chlorocebus aethiops
medicine
Animals
Humans
Receptors, Immunologic
Molecular Biology
Vero Cells
Caliciviridae Infections
Viral Vaccines
04 agricultural and veterinary sciences
Cell Biology
biology.organism_classification
Virology
Reverse genetics
Caliciviridae
030104 developmental biology
Viral replication
Capsid
Capsid Proteins
Rabbits
Subjects
Details
- ISSN :
- 1083351X
- Volume :
- 292
- Issue :
- 21
- Database :
- OpenAIRE
- Journal :
- The Journal of biological chemistry
- Accession number :
- edsair.doi.dedup.....242d262e6680254cb4aae63149b61038