title: Kaposi's sarcoma-associated herpesvirus LANA recruits the DNA polymerase clamp loader to mediate efficient replication and virus persistence.
abstract: Kaposi's sarcoma-associated herpesvirus (KSHV) latently infects tumor cells and persists as a multiple-copy, extrachromosomal, circular episome. To persist, the viral genome must replicate with each cell cycle. The KSHV latency-associated nuclear antigen (LANA) mediates viral DNA replication and persistence, but little is known regarding the underlying mechanisms. We find that LANA recruits replication factor C (RFC), the DNA polymerase clamp [proliferating cell nuclear antigen (PCNA)] loader, to drive DNA replication efficiently. Mutated LANA lacking RFC interaction was deficient for LANA-mediated DNA replication and episome persistence. RFC depletion had a negative impact on LANA's ability to replicate and maintain viral DNA in cells containing artificial KSHV episomes or in infected cells, leading to loss of virus. LANA substantially increased PCNA loading onto DNA in vitro and recruited RFC and PCNA to KSHV DNA in cells. These findings suggest that PCNA loading is a rate-limiting step in DNA replication that is incompatible with viral survival. LANA enhancement of PCNA loading permits efficient virus replication and persistence, revealing a previously unidentified mechanism for KSHV latency.
authors: Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115;
authors: Qiming
authors: Q
authors: Sun
authors: Department of Biology, Kyushu University, Fukuoka 812-8581, Japan; and.
authors: Toshiki
authors: T
authors: Tsurimoto
authors: Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115;
authors: Franceline
authors: F
authors: Juillard
authors: Proteomics Center, National Institute of Biological Sciences, Beijing 102206, People's Republic of China.
authors: Lin
authors: L
authors: Li
authors: Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115;
authors: Shijun
authors: S
authors: Li
authors: Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115;
authors: Erika
authors: E
authors: De León Vázquez
authors: Proteomics Center, National Institute of Biological Sciences, Beijing 102206, People's Republic of China.
authors: She
authors: S
authors: Chen
authors: Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115; kkaye@rics.bwh.harvard.edu.
authors: Kenneth
authors: K
authors: Kaye
mesh_terms: D000956:Antigens, Viral / Q000502:physiology*; D045744:Cell Line, Tumor; D004261:DNA Replication / Q000502:physiology; D004279:DNA, Viral / Q000096:biosynthesis / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000502:physiology; D055785:Gene Knockdown Techniques; D019288:Herpesvirus 8, Human / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology*; D054884:Host-Pathogen Interactions; D006801:Humans; D009687:Nuclear Proteins / Q000502:physiology*; D018809:Proliferating Cell Nuclear Antigen / Q000502:physiology; D051818:Replication Protein C / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D012514:Sarcoma, Kaposi / Q000503:physiopathology / Q000821:virology; D017735:Virus Latency / Q000502:physiology; D014779:Virus Replication / Q000502:physiology*
chemical_list: D000956:Antigens, Viral; D004279:DNA, Viral; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen; C081632:RFC1 protein, human; C109540:latency-associated nuclear antigen; D004259:DNA-Directed DNA Polymerase; D051818:Replication Protein C