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PMID: 8799130 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Chromosomal double-strand break repair in Ku80-deficient cells.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 93 ·No. 17 ·1996-08-20 ·Pages 8929-33

Liang F, Romanienko PJ, Weaver DT, Jeggo PA, Jasin M

Abstract

The x-ray sensitive hamster cell line xrs-6 is deficient in DNA double-strand break (DSB) repair and exhibits impaired V(D)J recombination. The molecular defect in this line is in the 80-kDa subunit of the Ku autoantigen, a protein that binds to DNA ends and recruits the DNA-dependent protein kinase to DNA. Using an I-SceI endonuclease expression system, chromosomal DSB repair was examined in xrs-6 and parental CHO-K1 cell lines. A DSB in chromosomal DNA increased the yield of recombinants several thousand-fold above background in both the xrs-6 and CHO-K1 cells, with recombinational repair of DSBs occurring in as many as 1 of 100 cells electroporated with the endonuclease expression vector. Thus, recombinational repair of chromosomal DSBs can occur at substantial levels in mammalian cells and it is not grossly affected in our assay by a deficiency of the Ku autoantigen. Rejoining of broken chromosome ends (end-joining) near the site of the DSB was also examined. In contrast to recombinational repair, end-joining was found to be severely impaired in the xrs-6 cells. Thus, the Ku protein appears to play a critical role in only one of the chromosomal DSB repair pathways.

MeSH Terms
Animals Antigens, Nuclear Base Sequence Chromosomes/metabolism Cricetinae DNA Helicases DNA Repair DNA-Binding Proteins/genetics Deoxyribonucleases, Type II Site-Specific/genetics,metabolism Gene Targeting Ku Autoantigen Molecular Sequence Data Nuclear Proteins/genetics Radiation Tolerance/physiology Recombination, Genetic Saccharomyces cerevisiae Proteins
Chemicals
Antigens, Nuclear DNA-Binding Proteins Nuclear Proteins Saccharomyces cerevisiae Proteins SCEI protein, S cerevisiae Deoxyribonucleases, Type II Site-Specific DNA Helicases XRCC5 protein, human Xrcc6 protein, human Ku Autoantigen
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Liang F
Program of Molecular Biology, Sloan-Kettering Institute, New York, NY 10021, USA.
Romanienko P J
Weaver D T
Jeggo P A
Jasin M
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.93.17.8929
Published
1996-08-20
Pages
8929-33
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC38571
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.93.17.8929 PMC Full Text View journal details
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