Variable diversity joining recombination: nonhairpin coding ends in thymocytes of SCID and wild-type mice.
Initiation of V(D)J recombination results in broken DNA molecules with blunt recombination signal ends and covalently sealed (hairpin) coding ends. In SCID mice, coding joint formation is severely impaired and hairpin coding ends accumulate as a result of a deficiency in the catalytic subunit of DNA-dependent protein kinase, an enzyme involved in the repair of DNA double-strand breaks. In this study, we report that not all SCID coding ends are hairpinned. We have detected open Jdelta1 and Ddelta2 coding ends at the TCRdelta locus in SCID thymocytes. Approximately 25% of 5'Ddelta2 coding ends were found to be open. Large deletions and abnormally long P nucleotide additions typical of SCID Ddelta2-Jdelta1 coding joints were not observed. Most Jdelta1 and Ddelta2 coding ends exhibited 3' overhangs, but at least 20% had unique 5' overhangs not previously detected in vivo. We suggest that the SCID DNA-dependent protein kinase deficiency not only reduces the efficiency of hairpin opening, but also may affect the specificity of hairpin nicking, as well as the efficiency of joining open coding ends.
Nakajima
Pamela B
PB
Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, PA 19111, USA. PB_Nakajima@fccc.edu
Bosma
Melvin J
MJ
D020413:3' Untranslated Regions / Q000276:immunology / Q000378:metabolism; D020121:5' Untranslated Regions / Q000276:immunology / Q000378:metabolism; D000818:Animals; D000916:Antibody Diversity / Q000235:genetics*; D051747:DNA-Activated Protein Kinase; D004268:DNA-Binding Proteins; D005260:Female; D005333:Fetus / Q000166:cytology / Q000276:immunology; D007144:Immunoglobulin J-Chains / Q000235:genetics* / Q000378:metabolism; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008813:Mice, Inbred ICR; D018345:Mice, Knockout; D016513:Mice, SCID; D016366:Open Reading Frames / Q000276:immunology; D016133:Polymerase Chain Reaction / Q000379:methods; D017346:Protein Serine-Threonine Kinases / Q000172:deficiency / Q000235:genetics; D011995:Recombination, Genetic / Q000276:immunology*; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000378:metabolism*; D013950:Thymus Gland / Q000166:cytology* / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D004268:DNA-Binding Proteins; D007144:Immunoglobulin J-Chains; D007135:Immunoglobulin Variable Region; D051747:DNA-Activated Protein Kinase; D017346:Protein Serine-Threonine Kinases