Reference - PMID:22718908 - DNA polymerization-independent functions of DNA polymerase epsilon in assembly and progression of the replisome in fission yeast.
Reference summary
- PubMed ID
- PMID:22718908
- Title
- DNA polymerization-independent functions of DNA polymerase epsilon in assembly and progression of the replisome in fission yeast.
- Authors
- Handa T, Kanke M, Takahashi TS, Nakagawa T, Masukata H
- Citation
- Mol Biol Cell 2012 Aug;23(16):3240-53
- Publication year
- 2012
- Abstract
- DNA polymerase epsilon (Pol ε) synthesizes the leading strands, following the CMG (Cdc45, Mcm2-7, and GINS [Go-Ichi-Nii-San]) helicase that translocates on the leading-strand template at eukaryotic replication forks. Although Pol ε is essential for the viability of fission and budding yeasts, the N-terminal polymerase domain of the catalytic subunit, Cdc20/Pol2, is dispensable for viability, leaving the following question: what is the essential role(s) of Pol ε? In this study, we investigated the essential roles of Pol ε using a temperature-sensitive mutant and a recently developed protein-depletion (off-aid) system in fission yeast. In cdc20-ct1 cells carrying mutations in the C-terminal domain of Cdc20, the CMG components, RPA, Pol α, and Pol δ were loaded onto replication origins, but Cdc45 did not translocate from the origins, suggesting that Pol ε is required for CMG helicase progression. In contrast, depletion of Cdc20 abolished the loading of GINS and Cdc45 onto origins, indicating that Pol ε is essential for assembly of the CMG complex. These results demonstrate that Pol ε plays essential roles in both the assembly and progression of CMG helicase.
