{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,14]],"date-time":"2026-04-14T23:24:56Z","timestamp":1776209096455,"version":"3.50.1"},"reference-count":41,"publisher":"American Society for Microbiology","issue":"3","license":[{"start":{"date-parts":[[2017,3,1]],"date-time":"2017-03-01T00:00:00Z","timestamp":1488326400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2017,3]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Zika virus (ZIKV) is an emerging human pathogen that is spreading rapidly through the Americas and has been linked to the development of microcephaly and to a dramatically increased number of Guillain-Barr\u00e9 syndrome cases. Currently, no vaccine or therapeutic options for the prevention or treatment of ZIKV infections exist. In the study described in this report, we expressed, purified, and characterized full-length nonstructural protein 5 (NS5) and the NS5 polymerase domain (NS5pol) of ZIKV RNA-dependent RNA polymerase. Using purified NS5, we developed an\n            <jats:italic>in vitro<\/jats:italic>\n            nonradioactive primer extension assay employing a fluorescently labeled primer-template pair. Both purified NS5 and NS5pol can carry out\n            <jats:italic>in vitro<\/jats:italic>\n            RNA-dependent RNA synthesis in this assay. Our results show that Mn\n            <jats:sup>2+<\/jats:sup>\n            is required for enzymatic activity, while Mg\n            <jats:sup>2+<\/jats:sup>\n            is not. We found that ZIKV NS5 can utilize single-stranded DNA but not double-stranded DNA as a template or a primer to synthesize RNA. The assay was used to compare the efficiency of incorporation of analog 5\u2032-triphosphates by the ZIKV polymerase and to calculate their discrimination versus that of natural ribonucleotide triphosphates (rNTPs). The 50% inhibitory concentrations for analog rNTPs were determined in an alternative nonradioactive coupled-enzyme assay. We determined that, in general, 2\u2032-C-methyl- and 2\u2032-C-ethynyl-substituted analog 5\u2032-triphosphates were efficiently incorporated by the ZIKV polymerase and were also efficient chain terminators. Derivatives of these molecules may serve as potential antiviral compounds to be developed to combat ZIKV infection. This report provides the first characterization of ZIKV polymerase and demonstrates the utility of\n            <jats:italic>in vitro<\/jats:italic>\n            polymerase assays in the identification of potential ZIKV inhibitors.\n          <\/jats:p>","DOI":"10.1128\/aac.01967-16","type":"journal-article","created":{"date-parts":[[2016,12,23]],"date-time":"2016-12-23T17:28:40Z","timestamp":1482514120000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":44,"title":["Analysis of Ribonucleotide 5\u2032-Triphosphate Analogs as Potential Inhibitors of Zika Virus RNA-Dependent RNA Polymerase by Using Nonradioactive Polymerase Assays"],"prefix":"10.1128","volume":"61","author":[{"given":"Gaofei","family":"Lu","sequence":"first","affiliation":[{"name":"Emory Institute for Drug Development (EIDD), Emory University, Atlanta, Georgia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gregory R.","family":"Bluemling","sequence":"additional","affiliation":[{"name":"Emory Institute for Drug Development (EIDD), Emory University, Atlanta, Georgia, USA"},{"name":"Drug Innovation Ventures at Emory (DRIVE), Atlanta, Georgia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paul","family":"Collop","sequence":"additional","affiliation":[{"name":"Emory Institute for Drug Development (EIDD), Emory University, Atlanta, Georgia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Hager","sequence":"additional","affiliation":[{"name":"Emory Institute for Drug Development (EIDD), Emory University, Atlanta, Georgia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Damien","family":"Kuiper","sequence":"additional","affiliation":[{"name":"Emory Institute for Drug Development (EIDD), Emory University, Atlanta, Georgia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bharat P.","family":"Gurale","sequence":"additional","affiliation":[{"name":"Emory Institute for Drug Development (EIDD), Emory University, Atlanta, Georgia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"George R.","family":"Painter","sequence":"additional","affiliation":[{"name":"Emory Institute for Drug Development (EIDD), Emory University, Atlanta, Georgia, USA"},{"name":"Drug Innovation Ventures at Emory (DRIVE), Atlanta, Georgia, USA"},{"name":"Department of Pharmacology, Emory University School of Medicine, Atlanta, Georgia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Abel","family":"De La Rosa","sequence":"additional","affiliation":[{"name":"Emory Institute for Drug Development (EIDD), Emory University, Atlanta, Georgia, USA"},{"name":"Drug Innovation Ventures at Emory (DRIVE), Atlanta, Georgia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexander A.","family":"Kolykhalov","sequence":"additional","affiliation":[{"name":"Emory Institute for Drug Development (EIDD), Emory University, Atlanta, Georgia, USA"},{"name":"Drug Innovation Ventures at Emory (DRIVE), Atlanta, Georgia, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/0035-9203(52)90042-4"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.2807\/1560-7917.ES2014.19.13.20751"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid2102.141363"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00072-15"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1600651"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMra1114525"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1136\/bmj.i657"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00705-006-0903-z"},{"key":"e_1_3_3_10_2","first-page":"712","volume-title":"Fields virology","author":"Lindenbach B","year":"2013","unstructured":"Lindenbach B MurrayC ThielH RiceC. 2013. 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