The dyad symmetry element of Epstein-Barr virus is a dominant but dispensable replication origin

Elisabeth Ott, Paolo Norio, Marion Ritzi, Carl L. Schildkraut, Aloys Schepers

Research output: Contribution to journalArticle

12 Citations (Scopus)

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.

Original languageEnglish (US)
Article numbere18609
JournalPLoS One
Volume6
Issue number5
DOIs
StatePublished - 2011

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replication origin
Human herpesvirus 4
Replication Origin
Human Herpesvirus 4
Viruses
Origin Recognition Complex
Genes
Genome
genome
plasmids
Plasmids
Cells
genomics
B-lymphocytes
binding sites
cell cycle
Cell Cycle
B-Lymphocytes
Binding Sites

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

The dyad symmetry element of Epstein-Barr virus is a dominant but dispensable replication origin. / Ott, Elisabeth; Norio, Paolo; Ritzi, Marion; Schildkraut, Carl L.; Schepers, Aloys.

In: PLoS One, Vol. 6, No. 5, e18609, 2011.

Research output: Contribution to journalArticle

Ott, Elisabeth ; Norio, Paolo ; Ritzi, Marion ; Schildkraut, Carl L. ; Schepers, Aloys. / The dyad symmetry element of Epstein-Barr virus is a dominant but dispensable replication origin. In: PLoS One. 2011 ; Vol. 6, No. 5.
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