title: The dyad symmetry element of Epstein-Barr virus is a dominant but dispensable replication origin.
abstract: OriP, the latent origin of Epstein-Barr virus (EBV), consists of two essential elements: the dyad symmetry (DS) and the family of repeats (FR). The function of these elements has been predominantly analyzed in plasmids transfected into transformed cells. Here, we examined the molecular functions of DS in its native genomic context and at an ectopic position in the mini-EBV episome. Mini-EBV plasmids contain 41% of the EBV genome including all information required for the proliferation of human B cells. Both FR and DS function independently of their genomic context. We show that DS is the most active origin of replication present in the mini-EBV genome regardless of its location, and it is characterized by the binding of the origin recognition complex (ORC) allowing subsequent replication initiation. Surprisingly, the integrity of oriP is not required for the formation of the pre-replicative complex (pre-RC) at or near DS. In addition we show that initiation events occurring at sites other than the DS are also limited to once per cell cycle and that they are ORC-dependent. The deletion of DS increases initiation from alternative origins, which are normally used very infrequently in the mini-EBV genome. The sequence-independent distribution of ORC-binding, pre-RC-assembly, and initiation patterns indicates that a large number of silent origins are present in the mini-EBV genome. We conclude that, in mini-EBV genomes lacking the DS element, the absence of a strong ORC binding site results in an increase of ORC binding at dispersed sites.
authors: Department of Gene Vectors, Helmholtz Zentrum München, München, Germany.
authors: Elisabeth
authors: E
authors: Ott
authors: Paolo
authors: P
authors: Norio
authors: Marion
authors: M
authors: Ritzi
authors: Carl
authors: C
authors: Schildkraut
authors: Aloys
authors: A
authors: Schepers
references: Proc Natl Acad Sci U S A. 2008 Jul 15;105(28):9639-44
references: 18621728
references: J Virol. 2000 May;74(10):4512-22
references: 10775587
references: J Biol Chem. 2000 Nov 10;275(45):34983-8
references: 10945994
references: J Cell Biochem. 2005 Feb 1;94(2):247-56
references: 15546145
references: Mol Cell Biol. 2000 Nov;20(22):8602-12
references: 11046155
references: Mol Cell. 2005 Nov 23;20(4):575-87
references: 16307921
references: Mol Cell Biol. 1993 May;13(5):2688-96
references: 8386315
references: Nucleic Acids Res. 2001 Feb 1;29(3):809-17
references: 11160905
references: Mol Cell Biol. 2003 Feb;23(3):804-14
references: 12529386
references: J Cell Sci. 2002 Jan 1;115(Pt 1):51-9
references: 11801723
references: J Virol. 2004 Nov;78(21):11487-505
references: 15479791
references: Nucleic Acids Res. 1992 Nov 25;20(22):5971-8
references: 1461730
references: J Biochem. 1998 Jan;123(1):175-81
references: 9504426
references: Chromosome Res. 2010 Jan;18(1):63-77
references: 19904620
references: J Virol. 2004 Nov;78(22):12308-19
references: 15507618
references: EMBO J. 2007 Oct 3;26(19):4252-62
references: 17853891
references: J Virol. 1990 May;64(5):2407-10
references: 2157896
references: Proc Natl Acad Sci U S A. 2001 Aug 28;98(18):10085-9
references: 11517328
references: Mol Cell Biol. 1991 Mar;11(3):1464-72
references: 1996103
references: Nat Rev Mol Cell Biol. 2004 Oct;5(10):848-55
references: 15459665
references: J Virol. 2003 Nov;77(21):11767-80
references: 14557661
references: J Virol. 1991 Jan;65(1):483-8
references: 1845903
references: J Virol. 2010 Mar;84(5):2533-46
references: 20032186
references: EMBO J. 2001 Aug 15;20(16):4588-602
references: 11500385
references: J Cell Sci. 2007 May 1;120(Pt 9):1529-39
references: 17405814
references: Mol Cell Biol. 2001 Feb;21(4):1098-110
references: 11158297
references: J Virol. 2000 Sep;74(18):8563-74
references: 10954558
references: Mol Cell Biol. 1995 May;15(5):2893-903
references: 7739569
references: Virology. 2004 Sep 1;326(2):317-28
references: 15302216
references: EMBO J. 2004 Jan 14;23(1):191-201
references: 14685267
references: EMBO J. 2001 Aug 1;20(15):4263-77
references: 11483529
references: Mol Cell Biol. 1995 Jan;15(1):173-8
references: 7799923
references: J Virol. 1995 Feb;69(2):1280-3
references: 7815506
references: Science. 2001 Dec 14;294(5550):2361-4
references: 11743204
references: Cell. 2001 Aug 10;106(3):287-96
references: 11509178
references: J Virol. 2002 Mar;76(5):2480-90
references: 11836426
references: J Cell Sci. 2003 Oct 1;116(Pt 19):3971-84
references: 12953058
references: Mol Cell Biol. 2006 Oct;26(20):7731-46
references: 16954389
references: Nat Rev Genet. 2010 Oct;11(10):673-84
references: 20811343
references: EMBO Rep. 2000 Aug;1(2):140-4
references: 11265753
references: Nucleic Acids Res. 1991 Sep 25;19(18):5053-8
references: 1656387
references: Bioessays. 2003 Feb;25(2):116-25
references: 12539237
references: Science. 1997 Feb 14;275(5302):993-5
references: 9020085
references: J Virol. 1998 Jun;72(6):4657-66
references: 9573229
references: Mol Cell Biol. 1985 Aug;5(8):1822-32
references: 3018528
references: J Virol. 1987 May;61(5):1743-6
references: 3033303
references: Nat Cell Biol. 2006 Dec;8(12):1313-6
references: 17139278
references: J Virol. 1994 Mar;68(3):1913-25
references: 8107251
references: Proc Natl Acad Sci U S A. 1984 Jun;81(12):3806-10
references: 6328526
references: J Virol. 1999 May;73(5):4385-92
references: 10196336
references: Science. 2001 Oct 5;294(5540):96-100
references: 11588251
references: Cell. 1989 Aug 11;58(3):527-35
references: 2547525
references: PLoS Biol. 2004 Jun;2(6):e152
references: 15208711
references: Mol Cell Biol. 1989 Mar;9(3):1026-33
references: 2542763