title: Genetic disruption of KSHV major latent nuclear antigen LANA enhances viral lytic transcriptional program.
abstract: Following primary infection, KSHV establishes a lifelong persistent latent infection in the host. The mechanism of KSHV latency is not fully understood. The latent nuclear antigen (LANA or LNA) encoded by ORF73 is one of a few viral genes expressed during KSHV latency, and is consistently detected in all KSHV-related malignancies. LANA is essential for KSHV episome persistence, and regulates the expression of viral lytic genes through epigenetic silencing, and inhibition of the expression and transactivation function of the key KSHV lytic replication initiator RTA (ORF50). In this study, we used a genetic approach to examine the role of LANA in regulating KSHV lytic replication program. Deletion of LANA did not affect the expression of its adjacent genes vCyclin (ORF72) and vFLIP (ORF71). In contrast, the expression levels of viral lytic genes including immediate-early gene RTA, early genes MTA (ORF57), vIL-6 (ORF-K2) and ORF59, and late gene ORF-K8.1 were increased before and after viral lytic induction with 12-O-tetradecanoyl-phorbol-13-acetate and sodium butyrate. This enhanced expression of viral lytic genes was also observed following overexpression of RTA with or without simultaneous chemical induction. Consistent with these results, the LANA mutant cells produced more infectious virions than the wild-type virus cells did. Furthermore, genetic repair of the mutant virus reverted the phenotypes to those of wild-type virus. Together, these results have demonstrated that, in the context of viral genome, LANA contributes to KSHV latency by regulating the expression of RTA and its downstream genes.
authors.affiliation: Tumor Virology Program, Greehey Children's Cancer Research Institute, Department of Microbiology and Immunology, The University of Texas Health Science Center at San Antonio, 8403 Floyd Curl Drive, San Antonio, TX 78229, USA.
authors.forename: Qiuhua
authors.initials: Q
authors.lastname: Li
authors.forename: Fuchun
authors.initials: F
authors.lastname: Zhou
authors.forename: Fengchun
authors.initials: F
authors.lastname: Ye
authors.forename: Shou-Jiang
authors.initials: SJ
authors.lastname: Gao
mesh_terms: D000956:Antigens, Viral / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D005814:Genes, Viral; D019288:Herpesvirus 8, Human / Q000235:genetics* / Q000276:immunology* / Q000472:pathogenicity / Q000502:physiology; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics* / Q000502:physiology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000235:genetics; D017735:Virus Latency / Q000235:genetics / Q000276:immunology; D014779:Virus Replication / Q000235:genetics / Q000276:immunology
chemical_list: D000956:Antigens, Viral; D017931:DNA Primers; D009687:Nuclear Proteins; D012333:RNA, Messenger; D012367:RNA, Viral; D014764:Viral Proteins; C109540:latency-associated nuclear antigen; C481275:viral FLIP protein, Human herpesvirus 8; D013755:Tetradecanoylphorbol Acetate