Methods for Studying Checkpoint Kinases – Chk1

Claudia Tapia-Alveal, Matthew J. O’Connell

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

3 Scopus citations

Abstract

Attempts to passage through mitosis with unrepaired DNA damage or incompletely replicated DNA leads to genome instability and/or cell death. To prevent this from occurring, an ancient checkpoint (known as the G2 DNA damage checkpoint) that inhibits the activation of the mitotic cyclin-dependent kinase is activated to hold cells in the G2 phase of the cell cycle. The effector of this checkpoint is Chk1, a protein serine-threonine kinase. Chk1 contains an N-terminal catalytic domain, and C-terminal regulatory domain. Within the regulatory domain there are two residues, Serine-317 (S317) and Serine-345 (S345), which are phosphorylated in active Chk1 molecules, and subsequently dephosphorylated to inactivate Chk1 and allow mitotic entry. Phospho-specific antibodies can be used to detect these activating phosphorylations, and this provides a simple and sensitive marker of Chk1 activation.

Original languageEnglish
Title of host publicationCell Cycle Checkpoints
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages171-179
Number of pages9
ISBN (Print)9781617792724
DOIs
StatePublished - 2011

Publication series

NameMethods in Molecular Biology
Volume782
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Chk1
  • DNA damage
  • Phosphorylation
  • Western blotting
  • checkpoint

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