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These virus-induced nuclear sites seem to behave as central hubs for the regulation of virus-host cell interactions, since proteins that promote efficient viral replication as well as factors that participate in the antiviral response are coopted and concentrated there. To gain further insight into the activities of viral RC, here we report, for the first time, the morphology, composition, and activities of RC isolated from Ad-infected cells. Morphological analyses of isolated RC particles by superresolution microscopy showed that they were indistinguishable from RC within infected cells and that they displayed a dynamic compartmentalization. Furthermore, the RC-containing fractions (RCf) proved to be functional, as they directed\n                    <jats:italic>de novo<\/jats:italic>\n                    synthesis of viral DNA and RNA as well as RNA splicing, activities that are associated with RC\n                    <jats:italic>in vivo<\/jats:italic>\n                    . A detailed analysis of the production of viral late mRNA from RCf at different times postinfection revealed that viral mRNA splicing occurs in RC and that the synthesis, posttranscriptional processing, and release from RC to the nucleoplasm of individual viral late transcripts are spatiotemporally separate events. The results presented here demonstrate that RCf are a powerful system for detailed study into RC structure, composition, and activities and, as a result, the determination of the molecular mechanisms that induce the formation of these viral sites of adenoviruses and other nuclear-replicating viruses.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>IMPORTANCE<\/jats:bold>\n                    RC may represent molecular hubs where many aspects of virus-host cell interaction are controlled. Here, we show by superresolution microscopy that RCf have morphologies similar to those of RC within Ad-infected cells and that they appear to be compartmentalized, as nucleolin and DBP display different localization in the periphery of these viral sites. RCf proved to be functional, as they direct\n                    <jats:italic>de novo<\/jats:italic>\n                    synthesis of viral DNA and mRNA, allowing the detailed study of the regulation of viral genome replication and expression. Furthermore, we show that the synthesis and splicing of individual viral late mRNA occurs in RC and that they are subject to different temporal patterns of regulation, from their synthesis to their splicing and release from RC to the nucleoplasm. Hence, RCf represent a novel system to study molecular mechanisms that are orchestrated in viral RC to take control of the infected cell and promote an efficient viral replication cycle.\n                  <\/jats:p>","DOI":"10.1128\/jvi.00033-16","type":"journal-article","created":{"date-parts":[[2016,1,14]],"date-time":"2016-01-14T04:42:57Z","timestamp":1452746577000},"page":"3411-3427","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":38,"title":["Morphological, Biochemical, and Functional Study of Viral Replication Compartments Isolated from Adenovirus-Infected Cells"],"prefix":"10.1128","volume":"90","author":[{"given":"Paloma","family":"Hidalgo","sequence":"first","affiliation":[{"name":"Centro de Investigaci\u00f3n en Din\u00e1mica Celular, Instituto de Investigaci\u00f3n en Ciencias B\u00e1sicas y Aplicadas, Universidad Aut\u00f3noma del Estado de Morelos (UAEM), Cuernavaca, Morelos, M\u00e9xico"},{"name":"Instituto de Biotecnolog\u00eda, Universidad Nacional Aut\u00f3noma de M\u00e9xico (UNAM), Cuernavaca, Morelos, M\u00e9xico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lourdes","family":"Anzures","sequence":"additional","affiliation":[{"name":"Centro de Investigaci\u00f3n en Din\u00e1mica Celular, Instituto de Investigaci\u00f3n en Ciencias B\u00e1sicas y Aplicadas, Universidad Aut\u00f3noma del Estado de Morelos (UAEM), Cuernavaca, Morelos, M\u00e9xico"},{"name":"Instituto de Biotecnolog\u00eda, Universidad Nacional Aut\u00f3noma de M\u00e9xico (UNAM), Cuernavaca, Morelos, M\u00e9xico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Armando","family":"Hern\u00e1ndez-Mendoza","sequence":"additional","affiliation":[{"name":"Centro de Investigaci\u00f3n en Din\u00e1mica Celular, Instituto de Investigaci\u00f3n en Ciencias B\u00e1sicas y Aplicadas, Universidad Aut\u00f3noma del Estado de Morelos (UAEM), Cuernavaca, Morelos, M\u00e9xico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ad\u00e1n","family":"Guerrero","sequence":"additional","affiliation":[{"name":"Laboratorio Nacional de Microscop\u00eda Avanzada, Instituto de Biotecnolog\u00eda, Universidad Nacional Aut\u00f3noma de M\u00e9xico (UNAM), Cuernavaca, Morelos, M\u00e9xico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christopher D.","family":"Wood","sequence":"additional","affiliation":[{"name":"Laboratorio Nacional de Microscop\u00eda Avanzada, Instituto de Biotecnolog\u00eda, Universidad Nacional Aut\u00f3noma de M\u00e9xico (UNAM), Cuernavaca, Morelos, M\u00e9xico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Margarita","family":"Vald\u00e9s","sequence":"additional","affiliation":[{"name":"Centro de Investigaci\u00f3n en Din\u00e1mica Celular, Instituto de Investigaci\u00f3n en Ciencias B\u00e1sicas y Aplicadas, Universidad Aut\u00f3noma del Estado de Morelos (UAEM), Cuernavaca, Morelos, M\u00e9xico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Dobner","sequence":"additional","affiliation":[{"name":"Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ram\u00f3n A.","family":"Gonzalez","sequence":"additional","affiliation":[{"name":"Centro de Investigaci\u00f3n en Din\u00e1mica Celular, Instituto de Investigaci\u00f3n en Ciencias B\u00e1sicas y Aplicadas, Universidad Aut\u00f3noma del Estado de Morelos (UAEM), Cuernavaca, Morelos, M\u00e9xico"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0065-3527(07)70004-0"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.coviro.2011.09.008"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.02046-13"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chom.2010.06.010"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1042\/BC20040058"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.5501\/wjv.v2.i2.32"},{"key":"e_1_3_2_8_2","first-page":"24388","article-title":"Architecture of the flaviviral replication complex","volume":"278","author":"Uchil PD","year":"2003","unstructured":"Uchil PD, Satchidanandam V. 2003. 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