Endonuclease activity inhibition of the NS1 protein of parvovirus B19 as a novel target for antiviral drug development

Peng Xu, Safder S. Ganaie, Xiaomei Wang, Zekun Wang, Steve Kleiboeker, Nancy C Horton, Richard F. Heier, Marvin J. Meyers, John E. Tavis, Jianming Qiu

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Human parvovirus B19 (B19V), a member of the genus Erythroparvovirus of the family Parvoviridae, is a small nonenveloped virus that has a single-stranded DNA (ssDNA) genome of 5.6 kb with two inverted terminal repeats (ITRs). B19V infection often results in severe hematological disorders and fetal death in humans. B19V replication follows a model of rolling hairpin-dependent DNA replication, in which the large nonstructural protein NS1 introduces a site-specific single-strand nick in the viral DNA replication origins, which locate at the ITRs. NS1 executes endonuclease activity through the N-terminal origin-binding domain. Nicking of the viral replication origin is a pivotal step in rolling hairpin-dependent viral DNA replication. Here, we developed a fluorophore-based in vitro nicking assay of the replication origin using the origin-binding domain of NS1 and compared it with the radioactive in vitro nicking assay. We used both assays to screen a set of small-molecule compounds (n 96) that have potential antinuclease activity. We found that the fluorophore-based in vitro nicking assay demonstrates sensitivity and specificity values as high as those of the radioactive assay. Among the 96 compounds, we identified 8 which have an inhibition of 80% at 10 M in both the fluorophore-based and radioactive in vitro nicking assays. We further tested 3 compounds that have a flavonoid-like structure and an in vitro 50% inhibitory concentration that fell in the range of 1 to 3 M. Importantly, they also exhibited inhibition of B19V DNA replication in UT7/Epo-S1 cells and ex vivo-expanded human erythroid progenitor cells.

Original languageEnglish (US)
Article numbere01879-18
JournalAntimicrobial Agents and Chemotherapy
DOIs
StatePublished - Mar 1 2019

Fingerprint

Endonucleases
Antiviral Agents
DNA Replication
Replication Origin
Terminal Repeat Sequences
Viral DNA
Parvoviridae
Human Parvovirus B19
Erythroid Precursor Cells
Fetal Death
Single-Stranded DNA
Flavonoids
Inhibitory Concentration 50
In Vitro Techniques
parvovirus NS1 protein
Genome
Viruses
Sensitivity and Specificity
Infection
Proteins

Keywords

  • Antivirals
  • In vitro nicking assay
  • Parvovirus B19

ASJC Scopus subject areas

  • Pharmacology
  • Pharmacology (medical)
  • Infectious Diseases

Cite this

Endonuclease activity inhibition of the NS1 protein of parvovirus B19 as a novel target for antiviral drug development. / Xu, Peng; Ganaie, Safder S.; Wang, Xiaomei; Wang, Zekun; Kleiboeker, Steve; Horton, Nancy C; Heier, Richard F.; Meyers, Marvin J.; Tavis, John E.; Qiu, Jianming.

In: Antimicrobial Agents and Chemotherapy, 01.03.2019.

Research output: Contribution to journalArticle

Xu, Peng ; Ganaie, Safder S. ; Wang, Xiaomei ; Wang, Zekun ; Kleiboeker, Steve ; Horton, Nancy C ; Heier, Richard F. ; Meyers, Marvin J. ; Tavis, John E. ; Qiu, Jianming. / Endonuclease activity inhibition of the NS1 protein of parvovirus B19 as a novel target for antiviral drug development. In: Antimicrobial Agents and Chemotherapy. 2019.
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AU - Kleiboeker, Steve

AU - Horton, Nancy C

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