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Each vRNA contains a central coding region that is flanked by noncoding regions. It has been shown that upon virion formation, the eight vRNAs are selectively packaged into progeny virions through segment\u2010specific packaging signals that are located in both the terminal coding regions and adjacent noncoding regions of each vRNA. Although recent studies using next\u2010generation sequencing suggest that multiple intersegment interactions are involved in genome packaging, contributions of the packaging signals to the intersegment interactions are not fully understood. Herein, using synthesized full\u2010length vRNAs of H1N1 WSN (A\/WSN\/33 [H1N1]) virus and short vRNAs containing the packaging signal sequences, we performed <jats:italic>in vitro<\/jats:italic> RNA binding assays and identified 15 intersegment interactions among eight vRNAs, most of which were mediated by the 3\u2032\u2010 and 5\u2032\u2010terminal regions. Interestingly, all eight vRNAs interacted with multiple other vRNAs, in that some bound to different vRNAs through their respective 3\u2032\u2010 and 5\u2032\u2010terminal regions. These <jats:italic>in vitro<\/jats:italic> findings would be of use in future studies of <jats:italic>in vivo<\/jats:italic> vRNA\u2013vRNA interactions during selective genome packaging.<\/jats:p>","DOI":"10.1111\/1348-0421.12766","type":"journal-article","created":{"date-parts":[[2019,12,16]],"date-time":"2019-12-16T10:32:26Z","timestamp":1576492346000},"page":"202-209","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["<i>In vitro<\/i> vRNA\u2013vRNA interactions in the H1N1 influenza A virus genome"],"prefix":"10.1111","volume":"64","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6541-1722","authenticated-orcid":false,"given":"Sho","family":"Miyamoto","sequence":"first","affiliation":[{"name":"Laboratory of Ultrastructural Virology, Institute for Frontier Life and Medical Sciences Kyoto University Kyoto Japan"},{"name":"Department of Molecular Virology, Graduate School of Medicine Kyoto University Kyoto Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0658-4663","authenticated-orcid":false,"given":"Takeshi","family":"Noda","sequence":"additional","affiliation":[{"name":"Laboratory of Ultrastructural Virology, Institute for Frontier Life and Medical Sciences Kyoto University Kyoto Japan"},{"name":"Laboratory of Ultrastructural Virology, Graduate School of Biostudies Kyoto University Kyoto Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2020,1,9]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkx584"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-02886-w"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41564-019-0513-7"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature04378"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1206069109"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.79.6.3766-3774.2005"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0437772100"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.80.5.2318-2325.2006"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01434-06"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02513-08"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.77.19.10575-10583.2003"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01301-13"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00662-12"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkm087"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01634-08"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2009.05.053"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.79.16.10348-10355.2005"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01541-07"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01144-07"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02267-07"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkr985"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gks1181"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.3390\/v8080218"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01022-14"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.3390\/v8080238"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2010.12.014"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.96.16.9345"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.79.11.6631-6643.2005"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-1896-6_12"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1038\/75125"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1261\/rna.678307"},{"key":"e_1_2_8_33_1","unstructured":"R Core Team. 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