Abstract
We recently have shown that mice deficient for the 86-kDa component (Ku80) of the DNA-dependent protein kinase exhibit growth retardation and a profound deficiency in V(D)J (variable, diversity, and joining) recombination. These defects may be related to abnormalities in DNA metabolism that arise from the inability of Ku80 mutant cells to process DNA double-strand breaks. To further characterize the role of Ku80 in DNA double-strand break repair, we have generated embryonic stem cells and pre-B cells and examined their response to ionizing radiation. Ku80(-/-) embryonic stem cells are more sensitive than controls to gamma-irradiation, and pre-B cells derived from Ku80 mutant mice display enhanced spontaneous and gamma-ray-induced apoptosis. We then determined the effects of ionizing radiation on the survival, growth, and lymphocyte development in Ku80-deficient mice. Ku80(-/-) mice display a hypersensitivity to gamma-irradiation, characterized by loss of hair pigmentation, severe injury to the gastrointestinal tract, and enhanced mortality. Exposure of newborn Ku80(-/-) mice to sublethal doses of ionizing radiation enhances their growth retardation and results in the induction of T cell-specific differentiation. However, unlike severe combined immunodeficient mice, radiation-induced T cell development in Ku80(-/-) mice is not accompanied by extensive thymocyte proliferation. The response of Ku80-deficient cell lines and mice to DNA-damaging agents provides important insights into the role of Ku80 in growth regulation, lymphocyte development, and DNA repair.
MeSH Terms
Animals
Antigens, Nuclear
B-Lymphocytes/cytology,immunology,radiation effects
Cell Differentiation/radiation effects
Cell Line
DNA Damage
DNA Helicases
DNA Repair
DNA-Binding Proteins/genetics,metabolism
Female
Gamma Rays/adverse effects
Growth/genetics,radiation effects
Growth Disorders/etiology,genetics
Intestines/pathology,radiation effects
Ku Autoantigen
Male
Mice
Mice, Knockout
Mice, SCID
Nuclear Proteins/genetics,metabolism
Radiation Tolerance
T-Lymphocytes/cytology,immunology,radiation effects
Chemicals
Antigens, Nuclear
DNA-Binding Proteins
Nuclear Proteins
DNA Helicases
XRCC5 protein, human
Xrcc6 protein, human
Xrcc6 protein, mouse
Ku Autoantigen
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Nussenzweig A
Departments of Medical Physics and Radiation Oncology, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.
Sokol K
Burgman P
Li L
Li G C
References (33)
33 references, click to expand
-
Restoration of X-ray resistance and V(D)J recombination in mutant cells by Ku cDNA.
Science. 1994 Oct 14;266(5183):288-91
PMID: 7939667
-
Sublethal gamma-radiation induces differentiation of CD4-/CD8- into CD4+/CD8+ thymocytes without T cell receptor beta rearrangement in recombinase activation gene 2-/- mice.
J Exp Med. 1994 Oct 1;180(4):1517-21
PMID: 7931082
-
DNA-dependent kinase (p350) as a candidate gene for the murine SCID defect.
Science. 1995 Feb 24;267(5201):1178-83
PMID: 7855601
-
Development of CD4+CD8+ thymocytes in RAG-deficient mice through a T cell receptor beta chain-independent pathway.
J Exp Med. 1995 Mar 1;181(3):1187-95
PMID: 7869035
-
Defective DNA-dependent protein kinase activity is linked to V(D)J recombination and DNA repair defects associated with the murine scid mutation.
Cell. 1995 Mar 10;80(5):813-23
PMID: 7889575
-
Loss of the catalytic subunit of the DNA-dependent protein kinase in DNA double-strand-break-repair mutant mammalian cells.
Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3171-4
PMID: 7724535
-
Isolation of mammalian cell mutants that are X-ray sensitive, impaired in DNA double-strand break repair and defective for V(D)J recombination.
Mutat Res. 1995 May;336(3):279-91
PMID: 7537861
-
What to do at an end: DNA double-strand-break repair.
Trends Genet. 1995 Oct;11(10):388-92
PMID: 7482764
-
Menage à trois: double strand break repair, V(D)J recombination and DNA-PK.
Bioessays. 1995 Nov;17(11):949-57
PMID: 8526889
-
Potential targets for autosomal SCID mutations.
Curr Opin Immunol. 1995 Aug;7(4):436-40
PMID: 7495505
-
The XRCC4 gene encodes a novel protein involved in DNA double-strand break repair and V(D)J recombination.
Cell. 1995 Dec 29;83(7):1079-89
PMID: 8548796
-
Requirement for Ku80 in growth and immunoglobulin V(D)J recombination.
Nature. 1996 Aug 8;382(6591):551-5
PMID: 8700231
-
Ku86-deficient mice exhibit severe combined immunodeficiency and defective processing of V(D)J recombination intermediates.
Cell. 1996 Aug 9;86(3):379-89
PMID: 8756720
-
Biochemical and genetic defects in the DNA-dependent protein kinase in murine scid lymphocytes.
Mol Cell Biol. 1996 Oct;16(10):5507-17
PMID: 8816463
-
Identification of a nonsense mutation in the carboxyl-terminal region of DNA-dependent protein kinase catalytic subunit in the scid mouse.
Proc Natl Acad Sci U S A. 1996 Sep 17;93(19):10285-90
PMID: 8816792
-
Evidence for DNA-PK-dependent and -independent DNA double-strand break repair pathways in mammalian cells as a function of the cell cycle.
Mol Cell Biol. 1997 Mar;17(3):1425-33
PMID: 9032269
-
Ku70-deficient embryonic stem cells have increased ionizing radiosensitivity, defective DNA end-binding activity, and inability to support V(D)J recombination.
Proc Natl Acad Sci U S A. 1997 Jul 22;94(15):8076-81
PMID: 9223317
-
Ku70 is required for DNA repair but not for T cell antigen receptor gene recombination In vivo.
J Exp Med. 1997 Sep 15;186(6):921-9
PMID: 9294146
-
A quantitative assay for transformation of bone marrow cells by Abelson murine leukemia virus.
J Exp Med. 1976 Jun 1;143(6):1453-63
PMID: 178822
-
Cell-cycle-dependent repair of potentially lethal damage in the XR-1 gamma-ray-sensitive Chinese hamster ovary cell.
Radiat Res. 1988 Aug;115(2):325-33
PMID: 3406371
-
Antibodies to CD3/T-cell receptor complex induce death by apoptosis in immature T cells in thymic cultures.
Nature. 1989 Jan 12;337(6203):181-4
PMID: 2521375
-
scid mutation in mice confers hypersensitivity to ionizing radiation and a deficiency in DNA double-strand break repair.
Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1394-7
PMID: 1996340
-
A link between double-strand break-related repair and V(D)J recombination: the scid mutation.
Proc Natl Acad Sci U S A. 1991 May 15;88(10):4061-5
PMID: 1709732
-
The SCID mouse mutant: definition, characterization, and potential uses.
Annu Rev Immunol. 1991;9:323-50
PMID: 1910681
-
Expression of the Ku protein during cell proliferation.
Biochim Biophys Acta. 1991 Oct 8;1090(2):181-7
PMID: 1932109
-
Features of apoptotic cells measured by flow cytometry.
Cytometry. 1992;13(8):795-808
PMID: 1333943
-
Impairment of V(D)J recombination in double-strand break repair mutants.
Science. 1993 Apr 9;260(5105):207-10
PMID: 8469973
-
Role of insulin-like growth factors in embryonic and postnatal growth.
Cell. 1993 Oct 8;75(1):73-82
PMID: 8402902
-
Chromosomal location and expression of the genes coding for Ku p70 and p80 in human cell lines and normal tissues.
Cytogenet Cell Genet. 1994;65(4):221-7
PMID: 8258294
-
DNA damage and the DNA-activated protein kinase.
Trends Biochem Sci. 1993 Nov;18(11):433-7
PMID: 8291090
-
Induction of T cell differentiation and lymphomagenesis in the thymus of mice with severe combined immune deficiency (SCID).
J Immunol. 1994 Aug 1;153(3):1004-14
PMID: 7913108
-
Phosphorylation of the p34 subunit of human single-stranded-DNA-binding protein in cyclin A-activated G1 extracts is catalyzed by cdk-cyclin A complex and DNA-dependent protein kinase.
Proc Natl Acad Sci U S A. 1994 Aug 30;91(18):8343-7
PMID: 8078885
-
Rescue of T cell-specific V(D)J recombination in SCID mice by DNA-damaging agents.
Science. 1994 Oct 21;266(5184):450-5
PMID: 7524150