Simple and rapid high-throughput assay to identify HSV-1 ICP0 transactivation inhibitors.

Autor: Ly CY; Department of Molecular Biosciences, The University of Kansas, Lawrence, KS, 66045, USA., Yu C; Department of Molecular Biosciences, The University of Kansas, Lawrence, KS, 66045, USA., McDonald PR; High Throughput Screening Laboratory, The University of Kansas, Lawrence, KS, 66047, USA., Roy A; High Throughput Screening Laboratory, The University of Kansas, Lawrence, KS, 66047, USA., Johnson DK; Computational Chemical Biology Core, The University of Kansas, Lawrence, KS, 66047, USA., Davido DJ; Department of Molecular Biosciences, The University of Kansas, Lawrence, KS, 66045, USA. Electronic address: ddavido@ku.edu.
Jazyk: angličtina
Zdroj: Antiviral research [Antiviral Res] 2021 Oct; Vol. 194, pp. 105160. Date of Electronic Publication: 2021 Aug 09.
DOI: 10.1016/j.antiviral.2021.105160
Abstrakt: Herpes simplex virus 1 (HSV-1) is a ubiquitous virus that results in lifelong infections due to its ability to cycle between lytic replication and latency. As an obligate intracellular pathogen, HSV-1 exploits host cellular factors to replicate and aid in its life cycle. HSV-1 expresses infected cell protein 0 (ICP0), an immediate-early regulator, to stimulate the transcription of all classes of viral genes via its E3 ubiquitin ligase activity. Here we report an automated, inexpensive, and rapid high-throughput approach to examine the effects of small molecule compounds on ICP0 transactivator function in cells. Two HSV-1 reporter viruses, KOS6β (wt) and dlx3.1-6β (ICP0-null mutant), were used to monitor ICP0 transactivation activity through the HSV-1 ICP6 promoter:lacz expression cassette. A ≥10-fold difference in β-galactosidase activity was observed in cells infected with KOS6β compared to dlx3.1-6β, demonstrating that ICP0 potently transactivates the ICP6 promoter. We established the robustness and reproducibility with a Z'-factor score of ≥0.69, an important criterium for high-throughput analyses. Approximately 19,000 structurally diverse compounds were screened and 76 potential inhibitors of the HSV-1 transactivator ICP0 were identified. We expect this assay will aid in the discovery of novel inhibitors and tools against HSV-1 ICP0. Using well-annotated compounds could identify potential novel factors and pathways that interact with ICP0 to promote HSV-1 gene expression.
(Copyright © 2021 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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