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Title: Defective chromatin recruitment and retention of NHEJ core components in human tumor cells expressing a Cyclin E fragment

Journal Article · · Nucleic Acids Research
DOI:https://doi.org/10.1093/nar/gkt812· OSTI ID:1625528
 [1];  [2];  [2];  [3];  [4];  [5]
  1. Cleveland Clinic, Cleveland, OH (United States). Lerner Research Inst. Dept. of Cancer Biology; Kent State Univ., Kent, OH (United States). School of Biomedical Sciences
  2. Cleveland Clinic, Cleveland, OH (United States). Lerner Research Inst. Dept. of Cancer Biology
  3. Cleveland Clinic, Cleveland, OH (United States). Lerner Research Inst. Dept. of Cancer Biology; Cleveland State Univ., Cleveland, OH (United States). Dept. of Chemistry
  4. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Life Sciences Division
  5. Cleveland Clinic, Cleveland, OH (United States). Lerner Research Inst. Dept. of Cancer Biology; Cleveland Clinic, Cleveland, OH (United States). Taussing Cancer Inst. Dept. of Radiation Oncology
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Exposure to genotoxic agents, such as ionizing radiation (IR), produces double-strand breaks, repaired predominantly in mammalian cells by non-homologous end-joining (NHEJ). Ku70 was identified as an interacting partner of a proteolytic Cyclin E (CycE) fragment, p18CycE. p18CycE endogenous generation during IR-induced apoptosis in leukemic cells and its stable expression in epithelial tumor cells sensitized to IR. cH2AX IR-induced foci (IRIFs) and comet assays indicated ineffective NHEJ DNA repair in p18CycE-expressing cells. DNA pull-down and chromatin recruitment assays revealed that retention of NHEJ factors to double strand breaks, but not recruitment, was diminished. Similarly, IRIFs of phosphorylated T2609 and S2056- DNA-PKcs and its target S1778-53BP1 were greatly decreased in p18CycE-expressing cells. As a result, DNA-PKcs chromatin association was also increased. 53BP1 IRIFs were suppressed when p18CycE was generated in leukemic cells and in epithelial cells stably expressing p18CycE. Ataxia telangiectasia mutated was activated but not its 53BP1 and MDC1 targets. These data indicate a profound influence of p18CycE on NHEJ through its interference with DNA-PKcs conformation and/ or dimerization, which is required for effective DNA repair, making the p18CycE-expressing cells more IR sensitive. These studies provide unique mechanistic insights into NHEJ misregulation in human tumor cells, in which defects in NHEJ core components are rare.

Research Organization:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division
Grant/Contract Number:
AC02-05CH11231
OSTI ID:
1625528
Journal Information:
Nucleic Acids Research, Vol. 41, Issue 22; ISSN 0305-1048
Publisher:
Oxford University PressCopyright Statement
Country of Publication:
United States
Language:
English

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Cited By (7)

PCNA-Ub polyubiquitination inhibits cell proliferation and induces cell-cycle checkpoints journal November 2016
Processing by MRE11 is involved in the sensitivity of subtelomeric regions to DNA double-strand breaks journal July 2015
PTEN Activation by DNA Damage Induces Protective Autophagy in Response to Cucurbitacin B in Hepatocellular Carcinoma Cells journal January 2016
Supraphysiological androgens suppress prostate cancer growth through androgen receptor–mediated DNA damage journal September 2019
PCNA-Ub polyubiquitination inhibits cell proliferation and induces cell-cycle checkpoints text January 2016
PCNA-Ub polyubiquitination inhibits cell proliferation and induces cell-cycle checkpoints text January 2016
USP14 is a deubiquitinase for Ku70 and critical determinant of non-homologous end joining repair in autophagy and PTEN-deficient cells journal November 2019

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