Chapter title |
PCNA-Dependent Ubiquitination of Cdt1 and p21 in Mammalian Cells.
|
---|---|
Chapter number | 19 |
Book title |
Cell Cycle Control
|
Published in |
Methods in molecular biology, January 2014
|
DOI | 10.1007/978-1-4939-0888-2_19 |
Pubmed ID | |
Book ISBNs |
978-1-4939-0887-5, 978-1-4939-0888-2
|
Authors |
Akiyo Hayashi, Naohiro Suenaga, Yasushi Shiomi, Hideo Nishitani, Hayashi, Akiyo, Suenaga, Naohiro, Shiomi, Yasushi, Nishitani, Hideo |
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 CRL4(Cdt2) for their degradation. Proteins degraded by CRL4(Cdt2) include Cdt1, p21, and Set8 in mammalian cells. These CRL4(Cdt2) 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 CRL4(Cdt2) 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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