Generation of an infectious clone of AMDV and identification of capsid residues essential for infectivity in cell culture

Virus Res. 2017 Oct 15:242:58-65. doi: 10.1016/j.virusres.2017.09.011. Epub 2017 Sep 18.

Abstract

Pathogenic strains of Aleutian mink disease virus (AMDV) such as Utah-1 do not replicate in cell culture (e.g., Crandell Rees feline kidney cells) while the in vitro-adapted AMDV strain ADV-Gorham (ADV-G) is not pathogenic. Here, we constructed a full-length infectious clone (pADV-G). Alignment of the VP2 gene of ADV-G with that of other AMDV strains revealed many amino acid (a.a.) residues conserved among pathogenic isolates that differed in ADV-G. Four virulence-associated, conserved residues of pADV-G VP2 were studied by site-directed mutagenesis (H92A, Q94S, Y115F, and I116L). Mutation of residue 92 or 94 decreased viral-transcription and viral-infectivity levels, whereas mutation of residue 115 or 116 did not affect viral-infectivity in CRFK cells. These results indicated that VP2 residues 92 and 94, both located on the surface of the viral capsid, are critical for AMDV infectivity in vitro.

Keywords: Aleutian mink disease virus; Functional residues; Infectious molecular clone; VP2 protein.

MeSH terms

  • Aleutian Mink Disease Virus / genetics
  • Aleutian Mink Disease Virus / physiology*
  • Animals
  • Capsid Proteins / genetics
  • Capsid Proteins / metabolism*
  • Cats
  • Cell Culture Techniques
  • Cells, Cultured
  • DNA Mutational Analysis
  • Mutagenesis, Site-Directed
  • Mutation, Missense
  • Reverse Genetics
  • Virus Internalization*
  • Virus Replication*

Substances

  • Capsid Proteins
  • VP2 protein, Aleutian mink disease virus