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(Received for publication, February 26, 1996)
From the Molecular Biology and the Cell Biology and Genetics
Programs, Sloan-Kettering Institute and Cornell University Graduate
School of Medical Sciences, New York, New York 10021
Mammalian cells possess a protein complex, termed
DNA-PK, which binds to DNA double strand breaks in vitro.
The complex consists of the heterodimeric Ku autoantigen and a
DNA-dependent protein kinase, DNA-PKcs. Cell
lines that are deficient for components of this complex are sensitive
to ionizing radiation and have impaired V(D)J recombination, a
site-specific recombination process. We have tested these cell lines
for their ability to repair double strand breaks in transfected DNA.
The xrs-6 cell line, which is deficient for the 80-kDa
subunit of the Ku autoantigen, exhibited reduced stability of
transfected DNA. Prior to obvious reductions in DNA stability, the
levels of homologous recombination and DNA end joining were unaffected.
However, the recovery of end joining products with precisely joined
ends was reduced, with a concomitant increase in products containing
deletions. Unlike the Ku80-deficient cells, no reduction in DNA
stability was detected in DNA-PKcs-deficient
scid cells. Scid cells also exhibited normal
levels of homologous recombination and DNA end joining. These
experiments implicate the Ku autoantigen, but not DNA-PKcs,
in a direct role in protecting DNA ends from degradation.
Volume 271, Number 24,
Issue of June 14, 1996
pp. 14405-14411
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
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