Regulation of Epstein-Barr virus OriP replication by poly(ADP-ribose) polymerase 1.
Poly(ADP-ribose) polymerase (PARP) is an abundant, chromatin-associated, NAD-dependent enzyme that functions in multiple chromosomal processes, including DNA replication and chromatin remodeling. The Epstein-Barr virus (EBV) origin of plasmid replication (OriP) is a dynamic genetic element that confers stable episome maintenance, DNA replication initiation, and chromatin organization functions. OriP function depends on the EBV-encoded origin binding protein EBNA1. We have previously shown that EBNA1 is subject to negative regulation by poly(ADP-ribosyl)ation (PARylation). We now show that PARP1 physically associates with OriP in latently EBV-infected B cells. Short hairpin RNA depletion of PARP1 enhances OriP replication activity and increases EBNA1, origin recognition complex 2 (ORC2), and minichromosome maintenance complex (MCM) association with OriP. Pharmacological inhibitors of PARP1 enhance OriP plasmid maintenance and increase EBNA1, ORC2, and MCM3 occupancy at OriP. PARylation in vitro inhibits ORC2 recruitment and remodels telomere repeat factor (TRF) binding at the dyad symmetry (DS) element of OriP. Purified PARP1 can ribosylate EBNA1 at multiple sites throughout its amino terminus but not in the carboxy-terminal DNA binding domain. We also show that EBNA1 linking regions (LR1 and LR2) can bind directly to oligomers of PAR. We propose that PARP1-dependent PARylation of EBNA1 and adjacently bound TRF2 induces structural changes at the DS element that reduce EBNA1 DNA binding affinity and functional recruitment of ORC.
Tempera
Italo
I
The Wistar Institute, 3601 Spruce Street, Philadelphia, PA 19104, USA.
Deng
Zhong
Z
Atanasiu
Constandache
C
Chen
Chi-Ju
CJ
D'Erme
Maria
M
Lieberman
Paul M
PM
D001402:B-Lymphocytes / Q000821:virology*; D002460:Cell Line; D019309:Epstein-Barr Virus Nuclear Antigens / Q000502:physiology*; D055785:Gene Knockdown Techniques; D004854:Herpesvirus 4, Human / Q000502:physiology*; D054884:Host-Pathogen Interactions; D006801:Humans; D010957:Plasmids; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases / Q000502:physiology*; D034741:RNA, Small Interfering / Q000235:genetics; D014779:Virus Replication
D019309:Epstein-Barr Virus Nuclear Antigens; D034741:RNA, Small Interfering; C499320:PARP1 protein, human; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases; C417029:EBV-encoded nuclear antigen 1