Human cytomegalovirus UL84 insertion mutant defective for viral DNA synthesis and growth

Journal of Virology
Yiyang XuGregory S Pari

Abstract

Human cytomegalovirus (HCMV) UL84 is required for oriLyt-dependent DNA replication, and evidence from transient transfection assays suggests that UL84 directly participates in DNA synthesis. In addition, because of its apparent interaction with IE2, UL84 is implicated as a possible regulatory protein. To address the role of UL84 in the context of the viral genome, we generated a recombinant HCMV bacterial artificial chromosome (BAC) construct that did not express the UL84 gene product. This construct, BAC-IN84/Ep, displayed a null phenotype in that it failed to produce infectious virus after transfection into human fibroblast cells, whereas a revertant virus readily produced viral plaques and, subsequently, infectious virus. Real-time quantitative PCR showed that BAC-IN84/Ep was defective for DNA synthesis in that no increase in the accumulation of viral DNA was observed in transfected cells. We were unable to complement BAC-IN84/Ep in trans; however, oriLyt-dependent DNA replication was observed by the cotransfection of UL84 and BAC-IN84/Ep. An analysis of viral mRNA by real-time PCR indicated that, even in the absence of DNA synthesis, all representative kinetic classes of genes were expressed in cells transfected with BAC-IN...Continue Reading

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Citations

Dec 6, 2006·Immunology and Cell Biology·Anthony A ScalzoMariapia A Degli-Esposti
Sep 29, 2005·Journal of Virology·Andrew J Davison, Nigel D Stow
Sep 22, 2010·Virology·David J Spector, Kristen Yetming
Sep 6, 2011·Virus Research·Li-Bo HeQi-Ya Zhang
Dec 18, 2013·Journal of Virology·Xiaofei ETimothy F Kowalik
Nov 21, 2007·Reviews in Medical Virology·Beatrice MercorelliGiorgio Palù

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