| Budget Amount *help |
¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000)
Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Outline of Final Research Achievements |
This research aimed to study the mechanism to proceed a replication fork with three functionally distinct DNA polymerases in eukaryotes. I especially focused on a biochemical background to distinguish DNA polymerases for syntheses of leading and lagging strands. Firstly, I reconstituted the replicative DNA helicase, human CMG complex and obtained the highly purified active CMG helicase. Secondly, I analyzed a functional significance of the interaction between the second PCNA loader Ctf18-RFC and DNA polymerase ε. I elucidated that Ctf18-RFC actively loads PCNA only when it complexes with DNA polymerase ε. This result indicated that Ctf18-RFC functions as a component of the leading-strand DNA polymerase complex and actively loads PCNA on the leading strands. Considering the role of RFC for loading of PCNA on the lagging strands, these two PCNA loaders will coordinate PCNA loading on both strands and play a role to distinguish DNA polymerases for syntheses of the two DNA strands.
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