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Nucleic Acids Research 2004 32(21):6218-6225; doi:10.1093/nar/gkh944
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Published online 1 December 2004

Nucleic Acids Research, Vol. 32 No. 21 © Oxford University Press 2004; all rights reserved

NFI and Oct-1 bend the Ad5 origin in the same direction leading to optimal DNA replication

Monika E. Mysiak, Claire Wyman1, P. Elly Holthuizen and Peter C. van der Vliet*

Department of Physiological Chemistry, University Medical Centre Utrecht and Centre for Biomedical Genetics, Universiteitsweg 100, 3584 CG Utrecht, The Netherlands and 1 Department of Radiation Oncology, Erasmus MC-Daniel and Department of Cell Biology and Genetics, Erasmus MC, PO Box 1738, 3000 DR, Rotterdam, The Netherlands

* To whom correspondence should be addressed. Tel: +31 302538973; Fax: +31 302539035; Email: p.c.vandervliet{at}med.uu.nl

Received August 27, 2004; Revised October 13, 2004; Accepted October 29, 2004

Two cellular transcription factors, nuclear factor I (NFI) and octamer binding protein (Oct-1), bind simultaneously to their recognition sequences in the Ad5 origin of replication thereby enhancing initiation. Using scanning force microscopy we have previously shown that NFI induces a 60° bend in the origin DNA. Here we demonstrate that Oct-1 induces a 42° bend in the origin DNA. Simultaneous binding of NFI and Oct-1 induces an 82° collective bend suggesting that both bends are oriented towards each other. In functional replication assays we further demonstrate that this extensive DNA bending leads to a synergistic enhancement of DNA replication. We propose that collective DNA bending induced by NFI and Oct-1 facilitates the optimal assembly of the preinitiation complex and plays an important role in the stimulatory mechanism of NFI and Oct-1 in replication.


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