Mapping of protein domains of hepatitis A virus 3AB essential for interaction with 3CD and viral RNA.
The small hydrophobic protein 3AB of the picornaviruses, encompassing the replication primer 3B, has been suggested to anchor the viral replication complex to membranes. For hepatitis A virus (HAV) 3AB, we have previously demonstrated its ability to form stable homodimers, to bind to membranes, and to interact specifically with RNA, implicating its multiple involvement in viral replication. In the present report, we show that HAV 3AB additionally interacts with HAV protein 3CD, a feature also described for the corresponding polypeptide of poliovirus. By assessing the interactions of three deletion mutants, distinct domains of HAV 3AB were mapped. The hydrophobic domain and the 3B moiety were found to be essential for the 3AB interaction with 3CD. Both electrostatic and hydrophobic forces are involved in this interaction. The cluster of charged amino acid residues at the C terminus of 3A seems to determine the specificity of 3AB interaction with RNA structures formed at either terminus of the HAV genome. Furthermore, our data implicate that 3A can interact with HAV RNA. Compared with poliovirus 3AB, which by itself is a nonspecific RNA-binding protein, HAV 3AB specifically recognizes HAV RNA structures that might be of relevance for initiation of viral RNA replication.
Beneduce
F
F
Laboratory of Virology, Istituto Superiore di Sanità, Viale Regina Elena 299, Rome, 00161, Italy.
Ciervo
A
A
Kusov
Y
Y
Gauss-Müller
V
V
Morace
G
G
D000086042:3C Viral Proteases; D000595:Amino Acid Sequence; D001665:Binding Sites; D002874:Chromosome Mapping; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D019281:Dimerization; D006507:Hepatovirus / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D014758:Viral Core Proteins / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins
D012367:RNA, Viral; D014758:Viral Core Proteins; D014764:Viral Proteins; D003546:Cysteine Endopeptidases; D000086042:3C Viral Proteases