Yeast Based Small Molecule Screen for Inhibitors of SARS-CoV Public Deposited

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Creator
  • Frieman, Matthew
    • Other Affiliation: University of Maryland
  • Basu, Dipanwita
    • Other Affiliation: University of Virginia School of Medicine
  • Matthews, Krystal
    • Other Affiliation: University of Maryland
  • Taylor, Justin
    • Other Affiliation: University of Maryland
  • Jones, Grant
    • Other Affiliation: University of Maryland
  • Pickles, Raymond
    • Affiliation: School of Medicine, Gene Therapy Center
  • Baric, Ralph S.
    • Affiliation: Gillings School of Global Public Health
  • Engel, Daniel A.
    • Other Affiliation: University of Virginia School of Medicine
Abstract
  • Severe acute respiratory coronavirus (SARS-CoV) emerged in 2002, resulting in roughly 8000 cases worldwide and 10% mortality. The animal reservoirs for SARS-CoV precursors still exist and the likelihood of future outbreaks in the human population is high. The SARS-CoV papain-like protease (PLP) is an attractive target for pharmaceutical development because it is essential for virus replication and is conserved among human coronaviruses. A yeast-based assay was established for PLP activity that relies on the ability of PLP to induce a pronounced slow-growth phenotype when expressed in S. cerevisiae. Induction of the slow-growth phenotype was shown to take place over a 60-hour time course, providing the basis for conducting a screen for small molecules that restore growth by inhibiting the function of PLP. Five chemical suppressors of the slow-growth phenotype were identified from the 2000 member NIH Diversity Set library. One of these, NSC158362, potently inhibited SARS-CoV replication in cell culture without toxic effects on cells, and it specifically inhibited SARS-CoV replication but not influenza virus replication. The effect of NSC158362 on PLP protease, deubiquitinase and anti-interferon activities was investigated but the compound did not alter these activities. Another suppressor, NSC158011, demonstrated the ability to inhibit PLP protease activity in a cell-based assay. The identification of these inhibitors demonstrated a strong functional connection between the PLP-based yeast assay, the inhibitory compounds, and SARS-CoV biology. Furthermore the data with NSC158362 suggest a novel mechanism for inhibition of SARS-CoV replication that may involve an unknown activity of PLP, or alternatively a direct effect on a cellular target that modifies or bypasses PLP function in yeast and mammalian cells.
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Resource type
  • Article
Rights statement
  • In Copyright
Journal title
  • PloS One
Journal volume
  • 6
Journal issue
  • 12
Language
  • English
ISSN
  • 1932-6203
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