Determinants in 3Dpol modulate the rate of growth of hepatitis A virus.
Hepatitis A virus (HAV), an atypical member of the Picornaviridae, grows poorly in cell culture. To define determinants of HAV growth, we introduced a blasticidin (Bsd) resistance gene into the virus genome and selected variants that grew at high concentrations of Bsd. The mutants grew fast and had increased rates of RNA replication and translation but did not produce significantly higher virus yields. Nucleotide sequence analysis and reverse genetic studies revealed that a T6069G change resulting in a F42L amino acid substitution in the viral polymerase (3D(pol)) was required for growth at high Bsd concentrations whereas a silent C7027T mutation enhanced the growth rate. Here, we identified a novel determinant(s) in 3D(pol) that controls the kinetics of HAV growth.
Konduru
Krishnamurthy
K
Laboratory of Hepatitis and Related Emerging Agents, DETTD-OBRR, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892, USA.
Kaplan
Gerardo G
GG
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D004252:DNA Mutational Analysis; D030041:Hepatitis A virus / Q000235:genetics* / Q000254:growth & development*; D008253:Macaca mulatta; D008958:Models, Molecular; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D017354:Point Mutation; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism*
C085421:3D protein, Hepatitis A virus; D014764:Viral Proteins