| Oxidative stress induces cell cycle-dependent Mre11 recruitment, ATM and Chk2 activation and histone H2AX phosphorylation. | |
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MedLine Citation:
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PMID: 18418078 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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DNA damage response recruits complex molecular machinery involved in DNA repair, arrest of cell cycle progression, and potentially in activation of apoptotic pathway. Among the first responders is the Mre11- (MRN) complex of proteins (Mre11, Rad50, Nbs1), essential for activation of ATM; the latter activates checkpoint kinase 2 (Chk2) and phosphorylates histone H2AX. In the present study the recruitment of Mre11 and phosphorylation of ATM, Chk2 and H2AX (gammaH2AX) detected immunocytochemically were measured by laser scanning cytometry to assess kinetics of these events in A549 cells treated with H(2)O(2). Recruitment of Mre11 was rapid, peaked at 10 min of exposure to the oxidant, and was of similar extent in all phases of the cell cycle. ATM and Chk2 activation as well as H2AX phosphorylation reached maximum levels after 30 min of treatment with H(2)O(2); the extent of phosphorylation of each was most prominent in S-, less in G(1)-, and the least in G(2)M- phase cells. A strong correlation between activation of ATM and Chk2, measured in the same cells, was seen in all phases of the cycle. In untreated cells activated Chk2 and Mre11 were distinctly present in centrosomes while in interphase cells they had characteristic punctate nuclear localization. The punctate expression of activated Chk2 both in untreated and H(2)O(2) treated cells was accentuated when measured as maximal pixel rather than integrated value of immunofluorescence (IF) per nucleus, and was most pronounced in G(1) cells, likely reflecting the function of Chk2 in activating Cdc25A. Subpopulations of G(1) and G(2)M cells with strong maximal pixel of Chk2-Thr68(P) IF in association with centrosomes were present in untreated cultures. Cytometric multiparameter assessment of the DNA damage response utilizing quantitative image analysis that allows one to measure inhomogeneity of fluorochrome distribution (e.g., maximal pixel) offers unique advantage in studies of the response of different cell constituents in relation to cell cycle position. |
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Authors:
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Hong Zhao; Frank Traganos; Anthony P Albino; Zbigniew Darzynkiewicz |
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Publication Detail:
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Type: Comparative Study; Journal Article; Research Support, N.I.H., Extramural Date: 2008-03-18 |
Journal Detail:
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Title: Cell cycle (Georgetown, Tex.) Volume: 7 ISSN: 1551-4005 ISO Abbreviation: Cell Cycle Publication Date: 2008 May |
Date Detail:
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Created Date: 2008-06-09 Completed Date: 2008-09-15 Revised Date: 2011-11-02 |
Medline Journal Info:
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Nlm Unique ID: 101137841 Medline TA: Cell Cycle Country: United States |
Other Details:
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Languages: eng Pagination: 1490-5 Citation Subset: IM |
Affiliation:
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Brander Cancer Research Institute and Department of Pathology, New York Medical College, Valhalla, New York 10595, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Cell Cycle
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physiology* Cell Cycle Proteins / metabolism* Cell Line, Tumor DNA Damage* DNA-Binding Proteins / metabolism* Enzyme Activation / physiology Histones / metabolism* Humans Hydrogen Peroxide Immunohistochemistry Kinetics Laser Scanning Cytometry Oxidative Stress / physiology* Phosphorylation Protein-Serine-Threonine Kinases / metabolism* Tumor Suppressor Proteins / metabolism* |
| Grant Support | |
ID/Acronym/Agency:
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CA 28704/CA/NCI NIH HHS; R01 CA028704-29/CA/NCI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Cell Cycle Proteins; 0/DNA-Binding Proteins; 0/H2AFX protein, human; 0/Histones; 0/MRE11A protein, human; 0/Tumor Suppressor Proteins; 7722-84-1/Hydrogen Peroxide; EC 2.7.1.11/checkpoint kinase 2; EC 2.7.11.1/Protein-Serine-Threonine Kinases; EC 2.7.11.1/ataxia telangiectasia mutated protein |
| Comments/Corrections | |
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