title: Detection of early Abl kinase activation after ionizing radiation by using a peptide biosensor.
abstract: The ubiquitously expressed Abl protein is a non-receptor tyrosine kinase that undergoes nuclear-cytoplasmic shuttling and is involved in many signaling pathways in the cell. Nuclear Abl is activated by DNA damage to regulate DNA repair, cell-cycle checkpoints and apoptosis. Previous studies have established that ataxia telangiectasia mutated (ATM) activates nuclear Abl by phosphorylating serine 465 (S465) in the kinase domain in response to ionizing radiation (IR). Using a peptide biosensor that specifically reports on the Abl kinase activity, we found that an Abl-S465A mutant, which is not capable of being activated by ATM through the canonical site, was still activated rapidly after IR. We established that DNA-dependent protein kinase (DNAPK) is likely to be responsible for a second pathway to activate Abl early on in the response to IR through phosphorylation at a site other than S465. Our findings show that nuclear and cytoplasmic Abl kinase is activated early on (within 5 min) in response to IR by both ATM and DNAPK, and that although one or the other of these kinases is required, either one is sufficient to activate Abl. These results support the concept of early Abl recruitment by both the ATM and the DNAPK pathways to regulate nuclear events triggered by DNA damage and potentially communicate them to proteins in the cytoplasm.
authors.affiliation: Department of Medicinal Chemistry and Molecular Pharmacology, College of Pharmacy, Center for Cancer Research, Purdue University, West Lafayette, IN 47907, USA.
authors.forename: Jiabin
authors.initials: J
authors.lastname: Tang
authors.forename: Jean Y
authors.initials: JY
authors.lastname: Wang
authors.forename: Laurie L
authors.initials: LL
authors.lastname: Parker
mesh_terms: D001549:Benzamides; D015374:Biosensing Techniques / Q000379:methods*; D002478:Cells, Cultured; D004249:DNA Damage; D004789:Enzyme Activation / Q000528:radiation effects; D057809:HEK293 Cells; D006801:Humans; D000068877:Imatinib Mesylate; D007259:Infrared Rays; D009025:Morpholines / Q000494:pharmacology; D010455:Peptides / Q000032:analysis* / Q000138:chemical synthesis / Q000737:chemistry; D010766:Phosphorylation; D010879:Piperazines / Q000494:pharmacology; D016315:Proto-Oncogene Proteins c-abl / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011743:Pyrimidines / Q000494:pharmacology; D011753:Pyrones / Q000494:pharmacology; D011839:Radiation, Ionizing; D012694:Serine / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: C495818:2-morpholin-4-yl-6-thianthren-1-yl-pyran-4-one; D001549:Benzamides; D009025:Morpholines; D010455:Peptides; D010879:Piperazines; D011743:Pyrimidines; D011753:Pyrones; D012694:Serine; D000068877:Imatinib Mesylate; D016315:Proto-Oncogene Proteins c-abl