title: Importance of amino acid 216 in nonstructural protein 2B for replication of hepatitis A virus in cell culture and in vivo.
abstract: Clinical isolates of hepatitis A virus (HAV) replicate inefficiently in cell culture unless mutations are acquired throughout the genome. An Ala-to-Val substitution in the nonstructural protein 2B (2B-216) was known to have a major impact on replication in cell culture. Analysis of chimeric viruses confirmed that the 2B-A[216]V change was critical for efficient replication and that Leu or Ile could substitute for Val. Viruses containing Val, Ile, or Leu at 2B-216 all replicated with similar kinetics in cell culture, whereas the virus containing Ala at this position grew 10- to 20-fold less efficiently. In contrast, in vivo, virus with either Ala or Val at 2B-216 replicated equally efficiently when tested in a chimpanzee and in tamarins, and each amino acid was stably maintained. Attempts to complement wild-type 2B in trans with adapted 2B provided by co-infection with a second viable HAV mutant failed to enhance replication of the virus containing the wild-type 2B sequence.
authors.affiliation: Molecular Hepatitis Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infections Diseases, National Institutes of Health, Bethesda, Maryland 20892-8009, USA. jgraff@niaid.nih.gov
authors.forename: Judith
authors.initials: J
authors.lastname: Graff
authors.forename: Suzanne U
authors.initials: SU
authors.lastname: Emerson
mesh_terms: D000818:Animals; D002460:Cell Line; D005814:Genes, Viral / Q000235:genetics; D005818:Genetic Engineering; D030041:Hepatitis A virus / Q000235:genetics* / Q000502:physiology*; D008253:Macaca mulatta / Q000821:virology; D009154:Mutation; D002679:Pan troglodytes / Q000821:virology; D010641:Phenotype; D012452:Saguinus / Q000821:virology; D012692:Serial Passage; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
chemical_list: C116507:2B protein, hepatitis A virus; D017361:Viral Nonstructural Proteins