Methods for studying checkpoint kinases - Chk1.
Methods for studying checkpoint kinases - Chk1.
复制标题
研究检查点激酶的方法 - Chk1。
DOI：
10.1007/978-1-61779-273-1_12
复制
发表时间：
2011
期刊：
影响因子：
--
通讯作者：
O'Connell,MatthewJ
中科院分区：
文献类型：
--
作者：
Tapia-Alveal,Claudia;O'Connell,MatthewJ
试图通过有丝分裂与未修复的DNA损伤或不完全复制的DNA导致基因组不稳定性和/或细胞死亡。为了防止这种情况发生，一个古老的检查点（称为G2 DNA损伤检查点），抑制有丝分裂细胞周期蛋白依赖性激酶的激活被激活，以保持细胞在细胞周期的G2期。该检查点的效应子是Chk 1，一种蛋白质丝氨酸-苏氨酸激酶。Chk 1含有N端催化结构域和C端调节结构域。在调节结构域内有两个残基，丝氨酸-317（S317）和丝氨酸-345（S345），其在活性Chk 1分子中被磷酸化，随后被去磷酸化以抑制Chk 1并允许有丝分裂进入。磷酸特异性抗体可用于检测这些激活磷酸化，这提供了一个简单而敏感的Chk 1激活标志物。
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.