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Analyzing DNA Replication Checkpoint in Budding Yeast

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Book cover Cell Cycle Control

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1170))

Abstract

Checkpoints are conserved mechanisms that prevent progression into the next phase of the cell cycle when cells are unable to accomplish the previous event properly. Cells also possess a surveillance mechanism called the DNA replication checkpoint, which consists of a conserved kinase cascade that is provoked by insults that block or slow down replication fork progression. In the budding yeast Saccharomyces cerevisiae, the DNA replication checkpoint controls the timing of S-phase events such as origin firing and spindle elongation. This checkpoint also upregulates dNTP pools and maintains the replication fork structure in order to resume DNA replication after replication block. Many replication checkpoint factors have been found to be tumor suppressors, highlighting the importance of this checkpoint pathway in human health. Here we describe a series of protocols to analyze the DNA replication checkpoint in S. cerevisiae.

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Acknowledgements

We thank Dr. Susan M. Gasser for critical reading and helpful suggestions. Support was gratefully received from the Novartis Research Foundation, the Swiss Cancer League, and the FP7 Marie Curie Network, Imaging the DNA Damage Response (Image DDR).

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Authors and Affiliations

  1. Friedrich Miescher Institute for Biomedical Research, Maulbeerstrasse 66, 4058, Basel, Switzerland

    Nicole Hustedt & Kenji Shimada

Authors
  1. Nicole Hustedt
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  2. Kenji Shimada
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Corresponding author

Correspondence to Kenji Shimada .

Editor information

Editors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Drexel University College of Medicine, Philadelphia, Pennsylvania, USA

    Eishi Noguchi

  2. Dept of Biochemistry & Molecular Biology, Drexel University College of Medicine, Philadelphia, USA

    Mariana C. Gadaleta

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Hustedt, N., Shimada, K. (2014). Analyzing DNA Replication Checkpoint in Budding Yeast. In: Noguchi, E., Gadaleta, M. (eds) Cell Cycle Control. Methods in Molecular Biology, vol 1170. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0888-2_16

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  • DOI: https://doi.org/10.1007/978-1-4939-0888-2_16

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0887-5

  • Online ISBN: 978-1-4939-0888-2

  • eBook Packages: Springer Protocols

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