VP1u phospholipase activity is critical for infectivity of full-length parvovirus B19 genomic clones

Claudia Filippone, Ning Zhi*, Susan Wong, Jun Lu, Sachiko Kajigaya, Giorgio Gallinella, Laura Kakkola, Maria Söderlund-Venermo, Neal S. Young, Kevin E. Brown

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    38 Citations (Scopus)

    Abstract

    Three full-length genomic clones (pB19-M20, pB19-FL and pB19-HG1) of parvovirus B19 were produced in different laboratories. pB19-M20 was shown to produce infectious virus. To determine the differences in infectivity, all three plasmids were tested by transfection and infection assays. All three clones were similar in viral DNA replication, RNA transcription, and viral capsid protein production. However, only pB19-M20 and pB19-HG1 produced infectious virus. Comparison of viral sequences showed no significant differences in ITR or NS regions. In the capsid region, there was a nucleotide sequence difference conferring an amino acid substitution (E176K) in the phospholipase A2-like motif of the VP1-unique (VP1u) region. The recombinant VP1u with the E176K mutation had no catalytic activity as compared with the wild-type. When this mutation was introduced into pB19-M20, infectivity was significantly attenuated, confirming the critical role of this motif. Investigation of the original serum from which pB19-FL was cloned confirmed that the phospholipase mutation was present in the native B19 virus.

    Original languageEnglish
    Pages (from-to)444-452
    Number of pages9
    JournalVirology
    Volume374
    Issue number2
    DOIs
    Publication statusPublished - 10 May 2008

    Keywords

    • Infectious clone
    • PLA-like motif
    • Parvovirus B19

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