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PMID: 7565721 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

scid cells are deficient in Ku and replication protein A phosphorylation by the DNA-dependent protein kinase.

Molecular and cellular biology ·Vol. 15 ·No. 10 ·1995-10-00 ·Pages 5700-6

Boubnov NV, Weaver DT

Abstract

Cell mutants of the Ku nuclear DNA-binding complex are ionizing radiation sensitive and show V(D)J recombination defects. Ku binds and activates a catalytic subunit of DNA-dependent protein kinase (DNA-PK), although the substrates for DNA-PK are unknown. We found that scid cell extracts were deficient in Ku phosphorylation by DNA-PK. Human chromosome 8-complemented scid cells, containing the human DNA-PK catalytic subunit, restored Ku phosphorylation. Likewise, radiation-induced RPA hyperphosphorylation was not completed in scid cells compared with control or chromosome 8-reconstituted cells. Thus, the inactivity of DNA-PK is likely responsible for the repair and recombination defects in scid cells.

MeSH Terms
Animals Antigens, Nuclear B-Lymphocytes Cell Extracts Cell Line Cell Line, Transformed Chromosomes, Human, Pair 8 DNA/metabolism DNA Helicases DNA Repair DNA-Activated Protein Kinase DNA-Binding Proteins/metabolism Fibroblasts Gamma Rays Genetic Complementation Test Humans Ku Autoantigen Mice Mice, SCID Nuclear Proteins/metabolism Phosphorylation/radiation effects Protein Serine-Threonine Kinases/metabolism Recombination, Genetic Replication Protein A Severe Combined Immunodeficiency/metabolism
Chemicals
Antigens, Nuclear Cell Extracts DNA-Binding Proteins Nuclear Proteins RPA1 protein, human Replication Protein A Rpa1 protein, mouse DNA DNA-Activated Protein Kinase PRKDC protein, human Protein Serine-Threonine Kinases DNA Helicases XRCC5 protein, human Xrcc6 protein, human Xrcc6 protein, mouse Ku Autoantigen
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Boubnov N V
Division of Tumor Immunology, Dana-Farber Cancer Institute, Boston, Massachusetts 02115, USA.
Weaver D T
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.15.10.5700
Published
1995-10-00
Pages
5700-6
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC230820
Subset
IM
Grants
NCI NIH HHS · CA54326 · United States
External Links
PubMed source DOI: 10.1128/MCB.15.10.5700 PMC Full Text View journal details
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