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Nucleic Acids Research Advance Access originally published online on October 11, 2007
Nucleic Acids Research 2007 35(20):6904-6916; doi:10.1093/nar/gkm822
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Nucleic Acids Research, 2007, Vol. 35, No. 20 6904-6916
© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Nucleic Acid Enzymes

Dynamics of human replication factors in the elongation phase of DNA replication

Yuji Masuda1,*, Miki Suzuki1, Jinlian Piao1, Yongqing Gu1, Toshiki Tsurimoto2 and Kenji Kamiya1

1Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima 734-8553, Japan and 2Department of Biology, School of Sciences, Kyushu University, Fukuoka 812-8581, Japan

* To whom correspondence should be addressed. Tel: +81 82 257 5893; Fax: +81 82 257 5843; Email: masudayu{at}hiroshima-u.ac.jp

Received July 19, 2007. Revised September 17, 2007. Accepted September 19, 2007.

In eukaryotic cells, DNA replication is carried out by coordinated actions of many proteins, including DNA polymerase {delta} (pol {delta}), replication factor C (RFC), proliferating cell nuclear antigen (PCNA) and replication protein A. Here we describe dynamic properties of these proteins in the elongation step on a single-stranded M13 template, providing evidence that pol {delta} has a distributive nature over the 7 kb of the M13 template, repeating a frequent dissociation–association cycle at growing 3'-hydroxyl ends. Some PCNA could remain at the primer terminus during this cycle, while the remainder slides out of the primer terminus or is unloaded once pol {delta} has dissociated. RFC remains around the primer terminus through the elongation phase, and could probably hold PCNA from which pol {delta} has detached, or reload PCNA from solution to restart DNA synthesis. Furthermore, we suggest that a subunit of pol {delta}, POLD3, plays a crucial role in the efficient recycling of PCNA during dissociation–association cycles of pol {delta}. Based on these observations, we propose a model for dynamic processes in elongation complexes.


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