TY - JOUR
T1 - Dynamics of human replication factors in the elongation phase of DNA replication
AU - Masuda, Yuji
AU - Suzuki, Miki
AU - Piao, Jinlian
AU - Gu, Yongqing
AU - Tsurimoto, Toshiki
AU - Kamiya, Kenji
N1 - Funding Information:
We thank Dr Marc S. Wold (University of Iowa College of Medicine, Iowa City, Iowa, USA) and Dr Tadashi Shimamoto (Hiroshima University, Hiroshima, Japan) for providing the RPA-expression plasmid and an E. coli strain to produce ss mp18 DNA, respectively. Several cloning vectors were obtained from the National BioResource Project (NIG, Japan). We thank Dr Haruo Ohmori (Institute for Virus Research, Kyoto University, Kyoto, Japan) and Dr Yoshihiro Matsumoto (Fox Chase Cancer Center, Philadelphia, USA) for helpful discussions. We are grateful to Eriko Aoki for her help with cDNA cloning, and Kumiko Mizuno, Tomoka Nakashima, Masako Okii, Hatsue Wakayama and Mai Yoshida for their laboratory assistance. This work was funded by Ministry of Education, Culture, Sports, Science and Technology of Japan (to Y.M., T.T., K.K.); the 21st Century Center of Excellence program from the Ministry of Education, Culture, Sports, Science and Technology of Japan (to K.K.); Research Fellowship for Young Scientists from Japan Society for the Promotion of Science (to J.P.); a scholarship award from the College Women’s Association of Japan (to Y.G.). Funding to pay the Open Access publication charges for this article was provided by Grants-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology of Japan (to K.K.).
PY - 2007/11
Y1 - 2007/11
N2 - In eukaryotic cells, DNA replication is carried out by coordinated actions of many proteins, including DNA polymerase δ (pol δ), 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 δ 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 δ has dissociated. RFC remains around the primer terminus through the elongation phase, and could probably hold PCNA from which pol δ has detached, or reload PCNA from solution to restart DNA synthesis. Furthermore, we suggest that a subunit of pol δ, POLD3, plays a crucial role in the efficient recycling of PCNA during dissociation-association cycles of pol δ. Based on these observations, we propose a model for dynamic processes in elongation complexes.
AB - In eukaryotic cells, DNA replication is carried out by coordinated actions of many proteins, including DNA polymerase δ (pol δ), 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 δ 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 δ has dissociated. RFC remains around the primer terminus through the elongation phase, and could probably hold PCNA from which pol δ has detached, or reload PCNA from solution to restart DNA synthesis. Furthermore, we suggest that a subunit of pol δ, POLD3, plays a crucial role in the efficient recycling of PCNA during dissociation-association cycles of pol δ. Based on these observations, we propose a model for dynamic processes in elongation complexes.
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U2 - 10.1093/nar/gkm822
DO - 10.1093/nar/gkm822
M3 - Article
C2 - 17932049
AN - SCOPUS:36749059976
SN - 0305-1048
VL - 35
SP - 6904
EP - 6916
JO - Nucleic acids research
JF - Nucleic acids research
IS - 20
ER -