ORC binding to TRF2 stimulates OriP replication.
In higher eukaryotes, the origin recognition complex (ORC) lacks sequence-specific DNA binding, and it remains unclear what other factors specify an origin of DNA replication. The Epstein-Barr virus origin of plasmid replication (OriP) recruits ORC, but the precise mechanism of ORC recruitment and origin activation is not clear. We now show that ORC is recruited selectively to the dyad symmetry (DS) region of OriP as a consequence of direct interactions with telomere repeat factor 2 (TRF2) and ORC1. TRF-binding sites within DS stimulate replication initiation and facilitate ORC recruitment in vitro and in vivo. TRF2, but not TRF1 or hRap1, recruits ORC from nuclear extracts. The amino-terminal domain of TRF2 associated with a specific region of ORC1 and was necessary for stimulation of DNA replication. These results support a model in which TRF2 stimulates OriP replication activity by direct binding with ORC subunits.
Atanasiu
Constandache
C
The Wistar Institute, 3601 Spruce Street, Philadelphia, Pennsylvania 19104, USA.
Deng
Zhong
Z
Wiedmer
Andreas
A
Norseen
Julie
J
Lieberman
Paul M
PM
D002460:Cell Line; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis / Q000235:genetics; D006367:HeLa Cells; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000378:metabolism; D006801:Humans; D009687:Nuclear Proteins / Q000378:metabolism*; D051738:Origin Recognition Complex / Q000378:metabolism*; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D018741:Replication Origin; D035182:TATA Box Binding Protein-Like Proteins / Q000378:metabolism*; D016615:Telomere / Q000235:genetics / Q000378:metabolism; D035341:Telomeric Repeat Binding Protein 2
D004279:DNA, Viral; D009687:Nuclear Proteins; D051738:Origin Recognition Complex; D011993:Recombinant Fusion Proteins; D035182:TATA Box Binding Protein-Like Proteins; C504753:TERF2 protein, human; D035341:Telomeric Repeat Binding Protein 2