Localization of MCM2-7, Cdc45, and GINS to the Site of DNA Unwinding during Eukaryotic DNA Replication
| Autor: | Yumiko Kubota, Johannes C. Walter, Antonin V. Tutter, Marcin Pacek, Haruhiko Takisawa |
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| Rok vydání: | 2006 |
| Předmět: |
DNA Replication
Macromolecular Substances DNA polymerase Xenopus DNA polymerase II Cell Cycle Proteins Eukaryotic DNA replication DNA-Directed DNA Polymerase Xenopus Proteins Control of chromosome duplication Animals Molecular Biology Adenosine Triphosphatases Genetics biology DNA replication Minichromosome Maintenance Complex Component 3 Nuclear Proteins Minichromosome Maintenance Complex Component 2 DNA Processivity Cell Biology Minichromosome Maintenance Complex Component 7 GINS biology.protein Nucleic Acid Conformation Replisome Carrier Proteins |
| Zdroj: | Molecular Cell. 21(4):581-587 |
| ISSN: | 1097-2765 |
| DOI: | 10.1016/j.molcel.2006.01.030 |
| Popis: | Little is known about the architecture and biochemical composition of the eukaryotic DNA replication fork. To study this problem, we used biotin-streptavidin-modified plasmids to induce sequence-specific replication fork pausing in Xenopus egg extracts. Chromatin immunoprecipitation was employed to identify factors associated with the paused fork. This approach identifies DNA pol alpha, DNA pol delta, DNA pol epsilon, MCM2-7, Cdc45, GINS, and Mcm10 as components of the vertebrate replisome. In the presence of the DNA polymerase inhibitor aphidicolin, which causes uncoupling of a highly processive DNA helicase from the stalled replisome, only Cdc45, GINS, and MCM2-7 are enriched at the pause site. The data suggest the existence of a large molecular machine, the "unwindosome," which separates DNA strands at the replication fork and contains Cdc45, GINS, and the MCM2-7 holocomplex. |
| Databáze: | OpenAIRE |
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