Analysis of the defect in DNA end joining in the murine scid mutation.
Murine severe combined immune deficiency (scid) is marked by a 5,000-fold reduction in coding joint formation in V(D)J recombination of antigen receptors. Others have demonstrated a sensitivity to double-strand breaks generated by ionizing radiation and bleomycin. We were interested in establishing the extent of the defect in intramolecular and intermolecular DNA end joining in lymphoid and nonlymphoid cells from scid mice. We conducted a series of studies probing the ability of these cells to resolve free ends of linear DNA molecules having various biochemical end configurations. We find that the stable integration of linear DNA into scid fibroblasts is reduced 11- to 75-fold compared with that in normal fibroblasts. In contrast, intramolecular and intermolecular end joining occur at normal frequencies in scid lymphocytes and fibroblasts. This normal level of end joining is observed regardless of the type of overhang and regardless of the requirement for nucleolytic activities prior to ligation. The fact that free ends having a wide variety of end configurations are recircularized normally in scid cells rules out certain models for the defect in scid. We discuss the types of DNA end joining reactions that are and are not affected in this double-strand break repair defect in the context of a hairpin model for V(D)J recombination.
Harrington
J
J
Department of Pathology, Stanford University School of Medicine, California 94305-5324.
Hsieh
C L
CL
Gerton
J
J
Bosma
G
G
Lieber
M R
MR
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004247:DNA; D004252:DNA Mutational Analysis; D004260:DNA Repair; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D015329:Gene Rearrangement, T-Lymphocyte; D016678:Genome; D008214:Lymphocytes / Q000166:cytology / Q000378:metabolism; D051379:Mice; D016513:Mice, SCID; D008969:Molecular Sequence Data; D009154:Mutation; D017385:Sequence Homology; D016511:Severe Combined Immunodeficiency / Q000235:genetics*
D004247:DNA