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.
CITATION STYLE
Tapia-Alveal, C., & O’Connell, M. J. (2011). Methods for Studying Checkpoint Kinases – Chk1. In Methods in Molecular Biology (Vol. 782, pp. 171–179). Humana Press Inc. https://doi.org/10.1007/978-1-61779-273-1_12
Mendeley helps you to discover research relevant for your work.