{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2022,4,13]],"date-time":"2022-04-13T05:07:51Z","timestamp":1649826471859},"reference-count":29,"publisher":"Portland Press Ltd.","issue":"3","content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2014,9,15]]},"abstract":"<jats:p>HIV-1 resistance to zidovudine [AZT (azidothymidine)] is associated with selection of the mutations M41L, D67N, K70R, L210W, T215F\/Y and K219Q\/E in RT (reverse transcriptase). These mutations decrease HIV-1 susceptibility to AZT by augmenting RT's ability to excise the chain-terminating AZT-MP (AZT-monophosphate) moiety from the chain-terminated DNA primer. Although AZT-MP excision occurs at the enzyme's polymerase active site, it is mechanistically distinct from the DNA polymerase reaction. Consequently, this activity represents a novel target for drug discovery, and inhibitors that target this activity may increase the efficacy of nucleoside\/nucleotide analogues, and may help to delay the onset of drug resistance. In the present study, we have developed a FRET (F\u00f6rster resonance energy transfer)-based high-throughput screening assay for the AZT-MP excision activity of RT. This assay is sensitive and robust, and demonstrates a signal-to-noise ratio of 3.3 and a Z\u2019 factor of 0.69. We screened three chemical libraries (7265 compounds) using this assay, and identified APEX57219 {3,3\u2032-[(3-carboxy-4-oxo-2,5-cyclohexadien-1-ylidene)methylene]bis[6-hydroxybenzoic acid]} as the most promising hit. APEX57219 displays a unique activity profile against wild-type and drug-resistant HIV-1 RT, and was found to inhibit virus replication at the level of reverse transcription. Mechanistic analyses revealed that APEX57219 blocked the interaction between RT and the nucleic acid substrate.<\/jats:p>","DOI":"10.1042\/bj20140365","type":"journal-article","created":{"date-parts":[[2014,6,27]],"date-time":"2014-06-27T12:15:12Z","timestamp":1403871312000},"page":"425-432","update-policy":"http:\/\/dx.doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Novel high-throughput screen identifies an HIV-1 reverse transcriptase inhibitor with a unique mechanism of action"],"prefix":"10.1042","volume":"462","author":[{"given":"Chih-Wei","family":"Sheen","sequence":"first","affiliation":[{"name":"Division of Infectious Diseases, Department of Medicine, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Onur","family":"Alpt\u00fcrk","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases, Department of Medicine, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicolas","family":"Sluis-Cremer","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases, Department of Medicine, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, U.S.A."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"288","published-online":{"date-parts":[[2014,8,22]]},"reference":[{"key":"2021112216063867900_B1","doi-asserted-by":"publisher","first-page":"40850","DOI":"10.1074\/jbc.R112.389056","article-title":"Human immunodeficiency virus reverse transcriptase: 25 years of research, drug discovery, and promise","volume":"287","author":"Le Grice","year":"2012","journal-title":"J. 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