Molecular and functional analyses of a human parvovirus B19 infectious clone demonstrates essential roles for NS1, VP1, and the 11-kilodalton protein in virus replication and infectivity.
In an attempt to experimentally define the roles of viral proteins encoded by the B19 genome in the viral life cycle, we utilized the B19 infectious clone constructed in our previous study to create two groups of B19 mutant genomes: (i) null mutants, in which either a translational initiation codon for each of these viral genes was substituted by a translational termination codon or a termination codon was inserted into the open reading frame by a frameshift; and (ii) a deletion mutant, in which half of the hairpin sequence was deleted at both the 5' and the 3' termini. The impact of these mutations on viral infectivity, DNA replication, capsid protein production, and distribution was systematically examined. Null mutants of the NS and VP1 proteins or deletion of the terminal hairpin sequence completely abolished the viral infectivity, whereas blocking expression of the 7.5-kDa protein or the putative protein X had no effect on infectivity in vitro. Blocking expression of the proline-rich 11-kDa protein significantly reduced B19 viral infectivity, and protein studies suggested that the expression of the 11-kDa protein was critical for VP2 capsid production and trafficking in infected cells. These findings suggest a previously unrecognized role for the 11-kDa protein, and together the results enhance our understanding of the key features of the B19 viral genome and proteins.
Zhi
Ning
N
Hematology Branch, National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892-1202, USA. zhin@nhlbi.nih.gov
Mills
Ian P
IP
Lu
Jun
J
Wong
Susan
S
Filippone
Claudia
C
Brown
Kevin E
KE
D036022:Capsid Proteins / Q000502:physiology*; D006801:Humans; D009154:Mutation; D010322:Parvoviridae Infections; D016732:Parvovirus B19, Human / Q000737:chemistry* / Q000472:pathogenicity / Q000502:physiology*; D017361:Viral Nonstructural Proteins / Q000502:physiology*; D014779:Virus Replication
C512342:11-kDa protein, human parvovirus B19; D036022:Capsid Proteins; D017361:Viral Nonstructural Proteins; C095547:capsid protein VP1, parvovirus B19