Adaptation of canine distemper virus to canine footpad keratinocytes modifies polymerase activity and fusogenicity through amino acid substitutions in the P/V/C and H proteins.
The wild-type canine distemper virus (CDV) strain A75/17 induces a non-cytocidal infection in cultures of canine footpad keratinocytes (CFKs) but produces very little progeny virus. After only three passages in CFKs, the virus produced 100-fold more progeny and induced a limited cytopathic effect. Sequence analysis of the CFK-adapted virus revealed only three amino acid differences, of which one was located in each the P/V/C, M and H proteins. In order to assess which amino acid changes were responsible for the increase of infectious virus production and altered phenotype of infection, we generated a series of recombinant viruses. Their analysis showed that the altered P/V/C proteins were responsible for the higher levels of virus progeny formation and that the amino acid change in the cytoplasmic tail of the H protein was the major determinant of cytopathogenicity.
Rivals
Jean-Paul
JP
Institut de Biotechnologie, Bâtiment de Biologie, University of Lausanne, CH-1015 Lausanne, Switzerland.
Plattet
Philippe
P
Currat-Zweifel
Christine
C
Zurbriggen
Andreas
A
Wittek
Riccardo
R
D000220:Adaptation, Biological / Q000235:genetics; D019943:Amino Acid Substitution; D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D003588:Cytopathogenic Effect, Viral / Q000235:genetics; D004252:DNA Mutational Analysis; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004217:Distemper Virus, Canine / Q000235:genetics / Q000254:growth & development*; D004285:Dogs; D015603:Keratinocytes / Q000821:virology*; D020125:Mutation, Missense; D011506:Proteins; D011995:Recombination, Genetic; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology
D011506:Proteins; D014764:Viral Proteins; D012321:DNA-Directed RNA Polymerases