title: The Kaposi's-sarcoma-associated herpesvirus orf35 gene product is required for efficient lytic virus reactivation.
abstract: Kaposi's sarcoma-associated herpesvirus (KSHV) is implicated in the etiology of several human malignancies. KSHV open reading frame (orf) 35 encodes a conserved gammaherpesvirus protein with an, as yet, unknown function. Employing the bacterial artificial chromosome (BAC) system, we generated a recombinant viral clone that fails to express ORF35 (BAC16-ORF35-stop) but preserves intact adjacent and overlapping reading frames. Using this construct, we studied the role of this previously uncharacterized gene product during lytic reactivation of KSHV. Upon lytic reactivation, the ORF35-stop recombinant virus displayed significantly reduced lytic viral gene expression, viral DNA replication, and progeny virus production as compared to control wild-type virus. Exogenous expression of ORF35-Flag reversed the effects of ORF35 deficiency. These results demonstrate that ORF35 is important for efficient lytic virus reactivation.
authors.affiliation: The Mina and Everard Goodman Faculty of Life Sciences and Advanced Materials and Nanotechnology Institute, Bar Ilan University, Ramat-Gan 5290002, Israel.
authors.forename: Shir
authors.initials: S
authors.lastname: Bergson
authors.affiliation: The Mina and Everard Goodman Faculty of Life Sciences and Advanced Materials and Nanotechnology Institute, Bar Ilan University, Ramat-Gan 5290002, Israel.
authors.forename: Inbal
authors.initials: I
authors.lastname: Itzhak
authors.affiliation: The Mina and Everard Goodman Faculty of Life Sciences and Advanced Materials and Nanotechnology Institute, Bar Ilan University, Ramat-Gan 5290002, Israel.
authors.forename: Talya
authors.initials: T
authors.lastname: Wasserman
authors.affiliation: The Mina and Everard Goodman Faculty of Life Sciences and Advanced Materials and Nanotechnology Institute, Bar Ilan University, Ramat-Gan 5290002, Israel.
authors.forename: Anastasia
authors.initials: A
authors.lastname: Gelgor
authors.affiliation: The Mina and Everard Goodman Faculty of Life Sciences and Advanced Materials and Nanotechnology Institute, Bar Ilan University, Ramat-Gan 5290002, Israel.
authors.forename: Inna
authors.initials: I
authors.lastname: Kalt
authors.affiliation: The Mina and Everard Goodman Faculty of Life Sciences and Advanced Materials and Nanotechnology Institute, Bar Ilan University, Ramat-Gan 5290002, Israel. Electronic address: saridr@mail.biu.ac.il.
authors.forename: Ronit
authors.initials: R
authors.lastname: Sarid
mesh_terms: D002460:Cell Line; D015870:Gene Expression; D015967:Gene Expression Regulation, Viral; D023061:Gene Order; D005816:Genetic Complementation Test; D005822:Genetic Vectors / Q000235:genetics; D019288:Herpesvirus 8, Human / Q000502:physiology*; D006801:Humans; D009154:Mutation; D016366:Open Reading Frames; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism; D014775:Virus Activation; D014779:Virus Replication
chemical_list: D014764:Viral Proteins
keywords: BAC16; Kaposi's sarcoma-associated herpesvirus, KSHV; Lytic infection; Open reading frame 35 (ORF35); Reactivation