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PCNA-Dependent Ubiquitination of Cdt1 and p21 in Mammalian Cells

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Book cover Cell Cycle Control

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

Abstract

PCNA is a DNA clamp, acting on chromatin as a platform for various proteins involved in many aspects of DNA replication-linked processes. Most of these proteins have the PCNA-interaction protein motif (PIP box) that associates with PCNA. Recent works show that PCNA plays an important role as a matchmaker, connecting PCNA-interacting proteins to the ubiquitin ligase CRL4Cdt2 for their degradation. Proteins degraded by CRL4Cdt2 include Cdt1, p21, and Set8 in mammalian cells. These CRL4Cdt2 substrates have a PIP degron that consists of the canonical PIP-box sequence and additional conserved amino acids required for ubiquitination. The degradation of these proteins is triggered when PCNA is loaded onto chromatin at the onset of S phase, and this process is important to prevent re-replication of DNA. These CRL4Cdt2 substrates are also degraded through the same mechanism in response to DNA damage. In this chapter, we describe several approaches to investigate how PIP degron-containing proteins are degraded in a PCNA-dependent manner.

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Acknowledgements

This work was financially supported by JSPS KAKENHI and MEXT KKENHI, Grant in Aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.

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Authors and Affiliations

  1. Graduate School of Life Science, University of Hyogo, Kamigori, Ako-gun, Hyogo, 678-1297, Japan

    Akiyo Hayashi, Naohiro Suenaga, Yasushi Shiomi & Hideo Nishitani

Authors
  1. Akiyo Hayashi
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  2. Naohiro Suenaga
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  3. Yasushi Shiomi
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  4. Hideo Nishitani
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Corresponding author

Correspondence to Hideo Nishitani .

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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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Hayashi, A., Suenaga, N., Shiomi, Y., Nishitani, H. (2014). PCNA-Dependent Ubiquitination of Cdt1 and p21 in Mammalian Cells. 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_19

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  • DOI: https://doi.org/10.1007/978-1-4939-0888-2_19

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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

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