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Article: High-resolution functional profiling of a gammaherpesvirus RTA locus in the context of the viral genome

TitleHigh-resolution functional profiling of a gammaherpesvirus RTA locus in the context of the viral genome
Authors
Issue Date2009
Citation
Journal of Virology, 2009, v. 83, n. 4, p. 1811-1822 How to Cite?
AbstractGammaherpesviruses Kaposi's sarcoma-associated herpesvirus and Epstein-Barr virus are associated with multiple human cancers. Our goal was to develop a quantitative, high-throughput functional profiling system to identify viral cis-elements and protein subdomains critical for virus replication in the context of the herpesvirus genome. In gamma-2 herpesviruses, the transactivating factor RTA is essential for initiation of lytic gene expression and viral reactivation. We used the RTA locus as a model to develop the functional profiling approach. The mutant murine gammaherpesvirus 68 viral library, containing 15-bp random insertions in the RTA locus, was passaged in murine fibroblast cells for multiple rounds of selection. The effect of each 15-bp insertion was characterized using fluorescent-PCR profiling. We identified 1,229 insertions in the 3,845-bp RTA locus, of which 393, 282, and 554 were critically impaired, attenuated, and tolerated, respectively, for viral growth. The functional profiling phenotypes were verified by examining several individual RTA mutant clones for transactivating function of the RTA promoter and transcomplementing function of the RTA-null virus. Thus, the profiling approach enabled us to identify several novel functional domains in the RTA locus in the context of the herpesvirus genome. Importantly, our study has demonstrated a novel system to conduct high-density functional genetic mapping. The genome-scale expansion of the genetic profiling approach will expedite the functional genomics research on herpesvirus. Copyright © 2009, American Society for Microbiology. All Rights Reserved.
Persistent Identifierhttp://hdl.handle.net/10722/285647
ISSN
2023 Impact Factor: 4.0
2023 SCImago Journal Rankings: 1.378
PubMed Central ID
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorArumugaswami, Vaithilingaraja-
dc.contributor.authorSitapara, Ronika-
dc.contributor.authorHwang, Seungmin-
dc.contributor.authorMoon, Jung Song-
dc.contributor.authorTuyet, Ngoc Ho-
dc.contributor.authorSu, Nancy Qi-
dc.contributor.authorSue, Eric Y.-
dc.contributor.authorKanagavel, Vidhya-
dc.contributor.authorXing, Fangfang-
dc.contributor.authorZhang, Xiaolin-
dc.contributor.authorZhao, Minglei-
dc.contributor.authorDeng, Hongyu-
dc.contributor.authorWu, Ting Ting-
dc.contributor.authorKanagavel, Sudhakar-
dc.contributor.authorZhang, Lu Lu-
dc.contributor.authorDandekar, Sugandha-
dc.contributor.authorPapp, Jeanette-
dc.contributor.authorSun, Ren-
dc.date.accessioned2020-08-18T04:56:17Z-
dc.date.available2020-08-18T04:56:17Z-
dc.date.issued2009-
dc.identifier.citationJournal of Virology, 2009, v. 83, n. 4, p. 1811-1822-
dc.identifier.issn0022-538X-
dc.identifier.urihttp://hdl.handle.net/10722/285647-
dc.description.abstractGammaherpesviruses Kaposi's sarcoma-associated herpesvirus and Epstein-Barr virus are associated with multiple human cancers. Our goal was to develop a quantitative, high-throughput functional profiling system to identify viral cis-elements and protein subdomains critical for virus replication in the context of the herpesvirus genome. In gamma-2 herpesviruses, the transactivating factor RTA is essential for initiation of lytic gene expression and viral reactivation. We used the RTA locus as a model to develop the functional profiling approach. The mutant murine gammaherpesvirus 68 viral library, containing 15-bp random insertions in the RTA locus, was passaged in murine fibroblast cells for multiple rounds of selection. The effect of each 15-bp insertion was characterized using fluorescent-PCR profiling. We identified 1,229 insertions in the 3,845-bp RTA locus, of which 393, 282, and 554 were critically impaired, attenuated, and tolerated, respectively, for viral growth. The functional profiling phenotypes were verified by examining several individual RTA mutant clones for transactivating function of the RTA promoter and transcomplementing function of the RTA-null virus. Thus, the profiling approach enabled us to identify several novel functional domains in the RTA locus in the context of the herpesvirus genome. Importantly, our study has demonstrated a novel system to conduct high-density functional genetic mapping. The genome-scale expansion of the genetic profiling approach will expedite the functional genomics research on herpesvirus. Copyright © 2009, American Society for Microbiology. All Rights Reserved.-
dc.languageeng-
dc.relation.ispartofJournal of Virology-
dc.titleHigh-resolution functional profiling of a gammaherpesvirus RTA locus in the context of the viral genome-
dc.typeArticle-
dc.description.naturelink_to_OA_fulltext-
dc.identifier.doi10.1128/JVI.02302-08-
dc.identifier.pmid19073723-
dc.identifier.pmcidPMC2643748-
dc.identifier.scopuseid_2-s2.0-59649086888-
dc.identifier.volume83-
dc.identifier.issue4-
dc.identifier.spage1811-
dc.identifier.epage1822-
dc.identifier.isiWOS:000262840200027-
dc.identifier.issnl0022-538X-

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