DNA polymerization-independent functions of DNA polymerase epsilon in assembly and progression of the replisome in fission yeast.

Tetsuya Handa, Mai Kanke, Tatsuro S. Takahashi, Takuro Nakagawa, Hisao Masukata

Research output: Contribution to journalArticle

23 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)3240-3253
Number of pages14
JournalMolecular biology of the cell
Volume23
Issue number16
Publication statusPublished - Jan 1 2012
Externally publishedYes

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DNA Polymerase II
Schizosaccharomyces
Polymerization
Saccharomycetales
Replication Origin
DNA
Catalytic Domain
Mutation
Temperature
Proteins

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

Cite this

DNA polymerization-independent functions of DNA polymerase epsilon in assembly and progression of the replisome in fission yeast. / Handa, Tetsuya; Kanke, Mai; Takahashi, Tatsuro S.; Nakagawa, Takuro; Masukata, Hisao.

In: Molecular biology of the cell, Vol. 23, No. 16, 01.01.2012, p. 3240-3253.

Research output: Contribution to journalArticle

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