{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T19:40:50Z","timestamp":1792093250624,"version":"4.4.0"},"reference-count":52,"publisher":"Oxford University Press (OUP)","issue":"2","license":[{"start":{"date-parts":[[2022,1,17]],"date-time":"2022-01-17T00:00:00Z","timestamp":1642377600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"name":"Singapore Ministry of Education Academic Research Fund Tier 2","award":["MOE-T2EP30220-0009"],"award-info":[{"award-number":["MOE-T2EP30220-0009"]}]},{"name":"Singapore Ministry of Education Academic Research Fund Tier 2","award":["MOE2016T22097"],"award-info":[{"award-number":["MOE2016T22097"]}]},{"DOI":"10.13039\/501100002301","name":"Estonian Research Council","doi-asserted-by":"publisher","award":["PRG1154"],"award-info":[{"award-number":["PRG1154"]}],"id":[{"id":"10.13039\/501100002301","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2022,1,25]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Alphaviruses such as Ross River virus (RRV), chikungunya virus (CHIKV), Sindbis virus (SINV), and Venezuelan equine encephalitis virus (VEEV) are mosquito-borne pathogens that can cause arthritis or encephalitis diseases. Nonstructural protein 4 (nsP4) of alphaviruses possesses RNA-dependent RNA polymerase (RdRp) activity essential for viral RNA replication. No 3D structure has been available for nsP4 of any alphaviruses despite its importance for understanding alphaviral RNA replication and for the design of antiviral drugs. Here, we report crystal structures of the RdRp domain of nsP4 from both RRV and SINV determined at resolutions of 2.6 \u00c5 and 1.9 \u00c5. The structure of the alphavirus RdRp domain appears most closely related to RdRps from pestiviruses, noroviruses, and picornaviruses. Hydrogen-deuterium exchange mass spectrometry (HDX-MS) and nuclear magnetic resonance (NMR) methods showed that in solution, nsP4 is highly dynamic with an intrinsically disordered N-terminal domain. Both full-length nsP4 and the RdRp domain were capable to catalyze RNA polymerization. Structure-guided mutagenesis using a trans-replicase system identified nsP4 regions critical for viral RNA replication.<\/jats:p>","DOI":"10.1093\/nar\/gkab1302","type":"journal-article","created":{"date-parts":[[2021,12,23]],"date-time":"2021-12-23T07:09:51Z","timestamp":1640243391000},"page":"1000-1016","source":"Crossref","is-referenced-by-count":53,"title":["Crystal structures of alphavirus nonstructural protein 4 (nsP4) reveal an intrinsically dynamic RNA-dependent RNA polymerase fold"],"prefix":"10.1093","volume":"50","author":[{"given":"Yaw Bia","family":"Tan","sequence":"first","affiliation":[{"name":"Lee Kong Chian School of Medicine, Nanyang Technological University, 59 Nanyang Drive, Singapore\u00a0636921"},{"name":"NTU Institute of Structural Biology, Nanyang Technological University, 59 Nanyang Drive, Singapore\u00a0636921"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laura\u00a0Sandra","family":"Lello","sequence":"additional","affiliation":[{"name":"University of Tartu, Institute of Technology, Nooruse 1, 50411 Tartu, Estonia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Liu","sequence":"additional","affiliation":[{"name":"Shanghai Institute of Materia Medica, China Academy of Sciences, 555 Zu Chong Zhi Road, Zhang Jiang Hi-Tech Park, Pudong, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yee-Song","family":"Law","sequence":"additional","affiliation":[{"name":"Lee Kong Chian School of Medicine, Nanyang Technological University, 59 Nanyang Drive, Singapore\u00a0636921"},{"name":"NTU Institute of Structural Biology, Nanyang Technological University, 59 Nanyang Drive, Singapore\u00a0636921"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Congbao","family":"Kang","sequence":"additional","affiliation":[{"name":"Experimental Drug Development Centre, Agency for Science, Technology and Research (A*STAR), 10 Biopolis Rd, #05-01\/06 Chromos, Singapore138670"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julien","family":"Lescar","sequence":"additional","affiliation":[{"name":"NTU Institute of Structural Biology, Nanyang Technological University, 59 Nanyang Drive, Singapore\u00a0636921"},{"name":"School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Zheng","sequence":"additional","affiliation":[{"name":"Shanghai Institute of Materia Medica, China Academy of Sciences, 555 Zu Chong Zhi Road, Zhang Jiang Hi-Tech Park, Pudong, Shanghai, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8193-0071","authenticated-orcid":false,"given":"Andres","family":"Merits","sequence":"additional","affiliation":[{"name":"University of Tartu, Institute of Technology, Nooruse 1, 50411 Tartu, Estonia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7637-7275","authenticated-orcid":false,"given":"Dahai","family":"Luo","sequence":"additional","affiliation":[{"name":"Lee Kong Chian School of Medicine, Nanyang Technological University, 59 Nanyang Drive, Singapore\u00a0636921"},{"name":"NTU Institute of Structural Biology, Nanyang Technological University, 59 Nanyang Drive, Singapore\u00a0636921"},{"name":"School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2022,1,17]]},"reference":[{"key":"2022012517583516700_B1","doi-asserted-by":"crossref","first-page":"e101","DOI":"10.1016\/S1473-3099(16)30518-7","article-title":"Epidemic arboviral diseases: priorities for research and public health","volume":"17","author":"Wilder-Smith","year":"2017","journal-title":"Lancet Infect. 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Virol."},{"key":"2022012517583516700_B6","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1186\/1471-2334-10-31","article-title":"Chronic pain associated with the chikungunya fever: long lasting burden of an acute illness","volume":"10","author":"de\u00a0Andrade","year":"2010","journal-title":"BMC Infect. Dis."},{"key":"2022012517583516700_B7","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1016\/j.virol.2017.02.014","article-title":"Discovery of a novel antiviral agent targeting the nonstructural protein 4 (nsP4) of chikungunya virus","volume":"505","author":"Wada","year":"2017","journal-title":"Virology"},{"key":"2022012517583516700_B8","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1128\/mr.58.3.491-562.1994","article-title":"The alphaviruses: gene expression, replication, and evolution","volume":"58","author":"Strauss","year":"1994","journal-title":"Microbiol. 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