Essential elements of a licensed, mammalian plasmid origin of DNA synthesis

Jindong Wang, Scott E. Lindner, Elizabeth R. Leight, Bill Sugden

27 Scopus citations

Abstract

We developed a mammalian plasmid replicon with a formerly uncharacterized origin of DNA synthesis, 8xRep*. 8xRep* functions efficiently to support once-per-cell-cycle synthesis of plasmid DNA which initiates within Rep*. By characterizing Rep*'s requirements for acting as an origin, we have uncovered several striking properties it shares with DS, the only other well-characterized, licensed, mammalian plasmid origin of DNA synthesis. Rep* contains a pair of previously unrecognized Epstein-Barr nuclear antigen 1 (EBNA1)-binding sites that are both necessary and sufficient in cis for its origin activity. These sites have an essential 21-bp center-to-center spacing, are bent by EBNA1, and recruit the origin recognition complex. The properties shared between DS and Rep* define cis and trans characteristics of a mammalian, extrachromosomal replicon. The role of EBNA1 likely reflects its evolution from cellular factors involved in the assembly of the initiation machinery.

Original languageEnglish (US)
Pages (from-to)1124-1134
Number of pages11
JournalMolecular and cellular biology
Volume26
Issue number3
DOIs
Publication statusPublished - Feb 1 2006

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

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