{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T00:18:34Z","timestamp":1790381914325,"version":"4.1.0"},"reference-count":48,"publisher":"American Society for Microbiology","issue":"19","license":[{"start":{"date-parts":[[2015,10,1]],"date-time":"2015-10-01T00:00:00Z","timestamp":1443657600000},"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":["J Virol"],"published-print":{"date-parts":[[2015,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Most new human infectious diseases emerge from cross-species pathogen transmissions; however, it is not clear how viruses adapt to productively infect new hosts. Host restriction factors represent one species-specific barrier that viruses may initially have little ability to inhibit in new hosts. For example, viral antagonists of protein kinase R (PKR) vary in their ability to block PKR-mediated inhibition of viral replication, in part due to PKR allelic variation between species. We previously reported that amplification of a weak PKR antagonist encoded by rhesus cytomegalovirus,\n            <jats:italic>rhtrs1<\/jats:italic>\n            , improved replication of a recombinant poxvirus (VV\u0394E\u0394K+RhTRS1) in several resistant primate cells. To test whether amplification increases the opportunity for mutations that improve virus replication with only a single copy of\n            <jats:italic>rhtrs1<\/jats:italic>\n            to evolve, we passaged\n            <jats:italic>rhtrs1<\/jats:italic>\n            -amplified viruses in semipermissive primate cells. After passage, we isolated two viruses that contained only a single copy of\n            <jats:italic>rhtrs1<\/jats:italic>\n            yet replicated as well as the amplified virus. Surprisingly,\n            <jats:italic>rhtrs1<\/jats:italic>\n            was not mutated in these viruses; instead, we identified mutations in two vaccinia virus (VACV) genes, A24R and A35R, either of which was sufficient to improve VV\u0394E\u0394K+RhTRS1 replication. Neither of these genes has previously been implicated in PKR antagonism. Furthermore, the mutation in A24R, but not A35R, increased resistance to the antipoxviral drug isatin-\u03b2-thiosemicarbazone, suggesting that these mutations employ different mechanisms to evade PKR. This study supports our hypothesis that gene amplification may provide a \u201cmolecular foothold,\u201d broadly improving replication to facilitate rapid adaptation, while subsequent mutations maintain this efficient replication in the new host without requiring gene amplification.\n          <\/jats:p>\n          <jats:p>\n            <jats:bold>IMPORTANCE<\/jats:bold>\n            Understanding how viruses adapt to a new host may help identify viruses poised to cross species barriers before an outbreak occurs. Amplification of\n            <jats:italic>rhtrs1<\/jats:italic>\n            , a weak viral antagonist of the host antiviral protein PKR, enabled a recombinant vaccinia virus to replicate in resistant cells from humans and other primates. After serial passage of\n            <jats:italic>rhtrs1<\/jats:italic>\n            -amplified viruses, there arose in two vaccinia virus genes mutations that improved viral replication without requiring\n            <jats:italic>rhtrs1<\/jats:italic>\n            amplification. Neither of these genes has previously been associated with inhibition of the PKR pathway. These data suggest that gene amplification can improve viral replication in a resistant host species and facilitate the emergence of novel adaptations that maintain the foothold needed for continued replication and spread in the new host.\n          <\/jats:p>","DOI":"10.1128\/jvi.01233-15","type":"journal-article","created":{"date-parts":[[2015,7,23]],"date-time":"2015-07-23T01:20:03Z","timestamp":1437614403000},"page":"9986-9997","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":32,"title":["Experimental Evolution Identifies Vaccinia Virus Mutations in A24R and A35R That Antagonize the Protein Kinase R Pathway and Accompany Collapse of an Extragenic Gene Amplification"],"prefix":"10.1128","volume":"89","author":[{"given":"Greg","family":"Brennan","sequence":"first","affiliation":[{"name":"Division of Human Biology, Fred Hutchinson Cancer Research Center, Seattle, Washington, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacob O.","family":"Kitzman","sequence":"additional","affiliation":[{"name":"Department of Human Genetics, University of Michigan, Ann Arbor, Michigan, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jay","family":"Shendure","sequence":"additional","affiliation":[{"name":"Department of Genome Sciences, University of Washington, Seattle, Washington, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adam P.","family":"Geballe","sequence":"additional","affiliation":[{"name":"Division of Human Biology, Fred Hutchinson Cancer Research Center, Seattle, Washington, USA"},{"name":"Division of Clinical Research, Fred Hutchinson Cancer Research Center, Seattle, Washington, USA"},{"name":"Departments of Microbiology and Medicine, University of Washington, Seattle, Washington, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1098\/rstb.2001.0888"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature06536"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1101\/cshperspect.a006841"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1404505"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.prevetmed.2012.11.021"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1007\/82_2012_223"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1038\/nri3295"},{"key":"e_1_3_2_9_2","first-page":"253","article-title":"Structure and function of the protein kinase R","volume":"316","author":"Sadler AJ","year":"2007","unstructured":"Sadler AJ, Williams BR. 2007. 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