Processing of 3'-Phosphoglycolate-Terminated DNA Double-StrandBreaks by Artemis Nuclease Metadata

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  • Main Title Processing of 3'-Phosphoglycolate-Terminated DNA Double-StrandBreaks by Artemis Nuclease


  • Author: Povrik, Lawrence F.
    Creator Type: Personal
  • Author: Zhou, Tong
    Creator Type: Personal
  • Author: Zhou, Ruizhe
    Creator Type: Personal
  • Author: Cowan, Morton J.
    Creator Type: Personal
  • Author: Yannone, Steven M.
    Creator Type: Personal


  • Sponsor: USDOE Director, Office of Science. Office of Biological andEnvironmental Research. Life Sciences Division
    Contributor Type: Organization


  • Name: Lawrence Berkeley National Laboratory
    Place of Publication: Berkeley, California
    Additional Info: "Ernest Orlando Lawrence Berkeley National Laboratory, Berkeley, CA (United States)"


  • Creation: 2005-10-01


  • English


  • Content Description: The Artemis nuclease is required for V(D)J recombination and for repair of an as yet undefined subset of radiation-induced DNA double-strand breaks. To assess the possibility that Artemis functions on oxidatively modified double-strand break termini, its activity toward model DNA substrates, bearing either 3{prime}-hydroxyl or 3{prime}-phosphoglycolate moieties, was examined. A 3{prime}-phosphoglycolate had little effect on Artemis-mediated trimming of long 3{prime} overhangs (>9 nucleotides), which were efficiently trimmed to 4-5 nucleotides. However, 3{prime}-phosphoglycolates on overhangs of 4-5 bases promoted selective Artemis-mediated trimming of a single 3{prime}-terminal nucleotide, while at least 2 nucleotides were trimmed from identical hydroxyl-terminated substrates. Artemis also efficiently removed a single nucleotide from a phosphoglycolate-terminated 3-base 3{prime} overhang, while leaving an analogous hydroxyl-terminated overhang largely intact. Such removal was dependent upon Ku, DNA-dependent protein kinase, and ATP. Together, these data suggest that Artemis-mediated cleavage of 3{prime} overhangs requires a minimum of 2 nucleotides, or a nucleotide plus a phosphoglycolate, 3{prime} to the cleavage site. Shorter 3{prime}-phosphoglycolate-terminated overhangs and blunt ends were also processed by Artemis, but much less efficiently. Consistent with the in vitro substrate specificity of Artemis, human cells lacking Artemis exhibited hypersensitivity to X-rays, bleomycin and neocarzinostatin, which all induce 3{prime}-phosphoglycolate-terminated double-strand breaks. Collectively, these results suggest that 3{prime}-phosphoglycolate termini and/or specific classes of DNA ends that arise from such blocked termini are relevant Artemis substrates in vivo.


  • Keyword: Bleomycin
  • Keyword: Nucleases
  • Keyword: Dna
  • Keyword: In Vitro
  • Keyword: Bearings
  • Keyword: Recombination
  • Keyword: In Vivo
  • Keyword: Proteins
  • STI Subject Categories: 60
  • Keyword: Specificity
  • Keyword: Animal Cells
  • Keyword: Repair
  • Keyword: Removal
  • Keyword: Cleavage
  • Keyword: Nucleotides
  • Keyword: Substrates


  • Journal Name: Journal of Biological Chemistry; Journal Volume: 282; Journal Issue: 6; Related Information: Journal Publication Date: 02/09/2007


  • Name: Office of Scientific & Technical Information Technical Reports
    Code: OSTI


  • Name: UNT Libraries Government Documents Department
    Code: UNTGD

Resource Type

  • Article


  • Text


  • Report No.: LBNL--59160
  • Grant Number: DE-AC02-05CH11231
  • Office of Scientific & Technical Information Report Number: 923183
  • Archival Resource Key: ark:/67531/metadc902296