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DNA polymerization-independent functions of DNA polymerase epsilon in assembly and progression of the replisome in fission yeast
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-...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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The American Society for Cell Biology
2012
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3418317/ https://www.ncbi.nlm.nih.gov/pubmed/22718908 http://dx.doi.org/10.1091/mbc.E12-05-0339 |
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author | Handa, Tetsuya Kanke, Mai Takahashi, Tatsuro S. Nakagawa, Takuro Masukata, Hisao |
author_facet | Handa, Tetsuya Kanke, Mai Takahashi, Tatsuro S. Nakagawa, Takuro Masukata, Hisao |
author_sort | Handa, Tetsuya |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-3418317 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-34183172012-10-30 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 Mol Biol Cell Articles 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. The American Society for Cell Biology 2012-08-15 /pmc/articles/PMC3418317/ /pubmed/22718908 http://dx.doi.org/10.1091/mbc.E12-05-0339 Text en © 2012 Handa et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society of Cell BD; are registered trademarks of The American Society of Cell Biology. |
spellingShingle | Articles Handa, Tetsuya Kanke, Mai Takahashi, Tatsuro S. Nakagawa, Takuro Masukata, Hisao DNA polymerization-independent functions of DNA polymerase epsilon in assembly and progression of the replisome in fission yeast |
title | DNA polymerization-independent functions of DNA polymerase epsilon in assembly and progression of the replisome in fission yeast |
title_full | DNA polymerization-independent functions of DNA polymerase epsilon in assembly and progression of the replisome in fission yeast |
title_fullStr | DNA polymerization-independent functions of DNA polymerase epsilon in assembly and progression of the replisome in fission yeast |
title_full_unstemmed | DNA polymerization-independent functions of DNA polymerase epsilon in assembly and progression of the replisome in fission yeast |
title_short | DNA polymerization-independent functions of DNA polymerase epsilon in assembly and progression of the replisome in fission yeast |
title_sort | dna polymerization-independent functions of dna polymerase epsilon in assembly and progression of the replisome in fission yeast |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3418317/ https://www.ncbi.nlm.nih.gov/pubmed/22718908 http://dx.doi.org/10.1091/mbc.E12-05-0339 |
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