Abstract
The checkpoint kinase Chk2 is activated in response to DNA damage through pathways requiring protein kinases ATM and/or ATR. The means by which Chk2 is activated by these kinases still remains to be addressed. Here we describe a cell-free system to study the activation of Chk2. Chk2 produced by a wheat germ extract in vitro transcription/translation system is inactive and can be activated by incubating with a rabbit reticulocyte lysate. This method will be useful for identification of cofactors required for activation of Chk2.
Publication types
-
Research Support, Non-U.S. Gov't
-
Research Support, U.S. Gov't, Non-P.H.S.
-
Research Support, U.S. Gov't, P.H.S.
MeSH terms
-
Animals
-
Ataxia Telangiectasia Mutated Proteins
-
Cell Cycle Proteins
-
Cell-Free System
-
Checkpoint Kinase 2
-
DNA Damage
-
DNA-Binding Proteins
-
Genetic Vectors
-
Humans
-
Immunoblotting
-
Plasmids / metabolism
-
Precipitin Tests
-
Protein Biosynthesis
-
Protein Serine-Threonine Kinases / metabolism*
-
Rabbits
-
Reticulocytes / metabolism
-
Transcription, Genetic
-
Triticum / metabolism
-
Tumor Suppressor Proteins
Substances
-
Cell Cycle Proteins
-
DNA-Binding Proteins
-
Tumor Suppressor Proteins
-
Checkpoint Kinase 2
-
ATM protein, human
-
Ataxia Telangiectasia Mutated Proteins
-
CHEK2 protein, human
-
Protein Serine-Threonine Kinases