title: Akt promotes post-irradiation survival of human tumor cells through initiation, progression, and termination of DNA-PKcs-dependent DNA double-strand break repair.
abstract: Akt phosphorylation has previously been described to be involved in mediating DNA damage repair through the nonhomologous end-joining (NHEJ) repair pathway. Yet the mechanism how Akt stimulates DNA-protein kinase catalytic subunit (DNA-PKcs)-dependent DNA double-strand break (DNA-DSB) repair has not been described so far. In the present study, we investigated the mechanism by which Akt can interact with DNA-PKcs and promote its function during the NHEJ repair process. The results obtained indicate a prominent role of Akt, especially Akt1 in the regulation of NHEJ mechanism for DNA-DSB repair. As shown by pull-down assay of DNA-PKcs, Akt1 through its C-terminal domain interacts with DNA-PKcs. After exposure of cells to ionizing radiation (IR), Akt1 and DNA-PKcs form a functional complex in a first initiating step of DNA-DSB repair. Thereafter, Akt plays a pivotal role in the recruitment of AKT1/DNA-PKcs complex to DNA duplex ends marked by Ku dimers. Moreover, in the formed complex, Akt1 promotes DNA-PKcs kinase activity, which is the necessary step for progression of DNA-DSB repair. Akt1-dependent DNA-PKcs kinase activity stimulates autophosphorylation of DNA-PKcs at S2056 that is needed for efficient DNA-DSB repair and the release of DNA-PKcs from the damage site. Thus, targeting of Akt results in radiosensitization of DNA-PKcs and Ku80 expressing, but not of cells deficient for, either of these proteins. The data showed indicate for the first time that Akt through an immediate complex formation with DNA-PKcs can stimulate the accumulation of DNA-PKcs at DNA-DSBs and promote DNA-PKcs activity for efficient NHEJ DNA-DSB repair.
authors.affiliation: Division of Radiobiology and Molecular Environmental Research, Department of Radiation Oncology, Eberhard Karls University Tübingen, Tübingen, Germany.
authors.forename: Mahmoud
authors.initials: M
authors.lastname: Toulany
authors.forename: Kyung-Jong
authors.initials: KJ
authors.lastname: Lee
authors.forename: Kazi R
authors.initials: KR
authors.lastname: Fattah
authors.forename: Yu-Fen
authors.initials: YF
authors.lastname: Lin
authors.forename: Brigit
authors.initials: B
authors.lastname: Fehrenbacher
authors.forename: Martin
authors.initials: M
authors.lastname: Schaller
authors.forename: Benjamin P
authors.initials: BP
authors.lastname: Chen
authors.forename: David J
authors.initials: DJ
authors.lastname: Chen
authors.forename: H Peter
authors.initials: HP
authors.lastname: Rodemann
mesh_terms: D002867:Chromones / Q000494:pharmacology; D053903:DNA Breaks, Double-Stranded / Q000187:drug effects / Q000528:radiation effects; D059766:DNA End-Joining Repair / Q000187:drug effects / Q000235:genetics; D004260:DNA Repair / Q000235:genetics; D051747:DNA-Activated Protein Kinase / Q000235:genetics / Q000378:metabolism; D045325:HCT116 Cells; D006801:Humans; D009025:Morpholines / Q000494:pharmacology; D046912:Multiprotein Complexes / Q000187:drug effects; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D011839:Radiation, Ionizing
chemical_list: C479235:2-(morpholin-4-yl)benzo(h)chromen-4-one; C499693:8-dibenzothiophen-4-yl-2-morpholin-4-yl-chromen-4-one; D002867:Chromones; D009025:Morpholines; D046912:Multiprotein Complexes; C494918:AKT1 protein, human; D051747:DNA-Activated Protein Kinase; D051057:Proto-Oncogene Proteins c-akt