title: ATM is the predominant kinase involved in the phosphorylation of histone H2AX after heating.
abstract: Heating induces histone H2AX phosphorylation at serine 139 (gammaH2AX). Phosphorylated H2AX subsequently forms foci in numerous mammalian cell lines. The aim of this study was to clarify details in the mechanisms involved in the phosphorylation of H2AX after heating. The cell lines used were DNA-PKcs knockout cells, ATM knockout cells, and their parental cell lines. To elucidate mechanisms of induction of phosphorylation of H2AX after heating, ATM/ATR inhibitor (CGK733) and DNA-PK inhibitor (NU7026) were used. The intensity of gammaH2AX signals was assayed with flow cytometry. The thermal dose-response curve for the fluorescence intensity of gammaH2AX appearance in DNA-PKcs-/- cells during the heating period was similar to that observed in DNA-PKcs+/+ cells. On the other hand, the slope of thermal dose-response curve for them in ATM-/- cells was lower than that in ATM+/+ cells. Phosphorylation of H2AX after heating was suppressed by a combination of CGK733 and NU7026 in the culture medium in DNA-PKcs-/- cells, ATM-/- cells and in their parental cells. Although the phosphorylation of H2AX after heating was not suppressed by NU7026 in their parental cells, such phosphorylation was suppressed by CGK733 in their parental cells. These results indicate that ATM is the predominant protein which is active in the phosphorylation of histone H2AX after heating.
authors.affiliation: Departments of Biology, School of Medicine, Nara Medical University, Nara, Japan. atakahas@naramed-u.ac.jp
authors.forename: Akihisa
authors.initials: A
authors.lastname: Takahashi
authors.forename: Eiichiro
authors.initials: E
authors.lastname: Mori
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Su
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Nakagawa
authors.forename: Noritomo
authors.initials: N
authors.lastname: Okamoto
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Uemura
authors.forename: Natsuko
authors.initials: N
authors.lastname: Kondo
authors.forename: Taichi
authors.initials: T
authors.lastname: Noda
authors.forename: Atsushi
authors.initials: A
authors.lastname: Toki
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Ejima
authors.forename: David J
authors.initials: DJ
authors.lastname: Chen
authors.forename: Ken
authors.initials: K
authors.lastname: Ohnishi
authors.forename: Takeo
authors.initials: T
authors.lastname: Ohnishi
mesh_terms: D000818:Animals; D064007:Ataxia Telangiectasia Mutated Proteins; D044482:Benzeneacetamides / Q000494:pharmacology; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002867:Chromones / Q000494:pharmacology; D051747:DNA-Activated Protein Kinase / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D006657:Histones / Q000737:chemistry / Q000378:metabolism*; D006358:Hot Temperature; D006801:Humans; D051379:Mice; D009025:Morpholines / Q000494:pharmacology; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation / Q000528:radiation effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D012694:Serine / Q000737:chemistry; D013890:Thiourea / Q000031:analogs & derivatives / Q000494:pharmacology; D025521:Tumor Suppressor Proteins / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*
chemical_list: D064007:Ataxia Telangiectasia Mutated Proteins; D044482:Benzeneacetamides; D018797:Cell Cycle Proteins; D002867:Chromones; D051747:DNA-Activated Protein Kinase; D004268:DNA-Binding Proteins; D006657:Histones; D009025:Morpholines; D009687:Nuclear Proteins; D047428:Protein Kinase Inhibitors; D017346:Protein Serine-Threonine Kinases; D012694:Serine; D013890:Thiourea; D025521:Tumor Suppressor Proteins; C479235:2-(morpholin-4-yl)benzo(h)chromen-4-one; C512273:CGK 733; C468783:H2AX protein, human; C576897:Atr protein, mouse; C576324:ATM protein, human; C576597:ATR protein, human; C576325:Atm protein, mouse; C497264:Prkdc protein, mouse; C475961:gamma-H2AX protein, mouse