A CRISPR/Cas9 Generated Bovine CD46-knockout Cell Line—A Tool to Elucidate the Adaptability of Bovine Viral Diarrhea Viruses (BVDV)
| Autor: | Dirk Höper, Susanne Koethe, Kerstin Wernike, Martin Beer, Kevin P Szillat |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Arginine
viruses Adaptation Biological lcsh:QR1-502 knockout ERNS escape mutant adaptation Virus Replication lcsh:Microbiology Madin Darby Canine Kidney Cells 0403 veterinary science Gene Knockout Techniques Viral Envelope Proteins Diarrhea Virus 2 Bovine Viral Asparagine Receptor CD46 MDBK Infectivity chemistry.chemical_classification 0303 health sciences biology Diarrhea Virus 1 Bovine Viral 04 agricultural and veterinary sciences 3. Good health Amino acid Infectious Diseases Ribonucleoproteins CRISPR Host-Pathogen Interactions Receptors Virus bovine viral diarrhea virus (BVDV) 040301 veterinary sciences Virus Article Membrane Cofactor Protein 03 medical and health sciences Dogs Virology Animals cell entry 030304 developmental biology pestivirus Pestivirus Virus Internalization biology.organism_classification chemistry Amino Acid Substitution Cell culture Cattle CRISPR-Cas Systems Protein Multimerization |
| Zdroj: | Viruses, Vol 12, Iss 859, p 859 (2020) Viruses Volume 12 Issue 8 |
| ISSN: | 1999-4915 |
| Popis: | Bovine viral diarrhea virus (BVDV) entry into a host cell is mediated by the interaction of the viral glycoprotein E2 with the cellular transmembrane CD46 receptor. In this study, we generated a stable Madin&ndash Darby Bovine Kidney (MDBK) CD46-knockout cell line to study the ability of different pestivirus A and B species (BVDV-1 and -2) to escape CD46-dependent cell entry. Four different BVDV-1/2 isolates showed a clearly reduced infection rate after inoculation of the knockout cells. However, after further passaging starting from the remaining virus foci on the knockout cell line, all tested virus isolates were able to escape CD46-dependency and grew despite the lack of the entry receptor. Whole-genome sequencing of the escape-isolates suggests that the genetic basis for the observed shift in infectivity is an amino acid substitution of an uncharged (glycine/asparagine) for a charged amino acid (arginine/lysine) at position 479 in the ERNS in three of the four isolates tested. In the fourth isolate, the exchange of a cysteine at position 441 in the ERNS resulted in a loss of ERNS dimerization that is likely to influence viral cell-to-cell spread. In general, the CD46-knockout cell line is a useful tool to analyze the role of CD46 for pestivirus replication and the virus&ndash receptor interaction. |
| Databáze: | OpenAIRE |
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