The scid factor on human chromosome 8 restores V(D)J recombination in addition to double-strand break repair.
The murine severe combined immune deficiency mutation (scid) is characterized by a lack of B- and T-lymphoid cells due to a defect in lymphoid V(D)J recombination. Moreover, defective rejoining of DNA double-strand breaks (dsb) in scid cells also results in a marked increase in sensitivity to ionizing radiation. Recently, the putative human homologue of the murine scid gene locus, HYRC1, was assigned to human chromosome 8q11, based on the radiation sensitivity of scid cells as compared to scid:human cell hybrids carrying portions of human chromosome 8. Given the precedent (e.g., ataxia-telangiectasia) for genes other than the affected one being able to complement radiation defects, we were interested in determining if the V(D)J recombination defect was also corrected by the HYRC1 locus. The V(D)J recombination analysis using extrachromosomal DNA substrates in control scid cells (SC3VA2) versus complemented cells (RD13B2) indicates that the radiation sensitivity-complemented cells (RD13B2) are also fully complemented for the V(D)J recombination reaction, whereas the control (uncomplemented) cells (SC3VA2) fail to carry out V(D)J recombination normally. Slightly over 60% of the radiation-induced dsb are rejoined even in scid cells, and this alternative pathway is temperature sensitive. Only the remaining 30-35% of dsb require the introduction of the HYRC1 locus, and this pathway is not temperature sensitive. This merely partial contribution of the scid factor to the repair process suggests the presence of another pathway of dsb repair. Our results indicate that the HYRC1 locus, assigned to human chromosome 8q11, encodes the scid factor, which is involved in all V(D)J recombination coding joint formation and in 30-35% of dsb repair by the temperature-resistant pathway.
Komatsu
K
K
Department of Radiation Biology, Hiroshima University, Japan.
Kubota
N
N
Gallo
M
M
Okumura
Y
Y
Lieber
M R
MR
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology; D002470:Cell Survival / Q000528:radiation effects; D002898:Chromosomes, Human, Pair 8; D004254:DNA Nucleotidyltransferases / Q000096:biosynthesis / Q000235:genetics*; D004260:DNA Repair; D004307:Dose-Response Relationship, Radiation; D005816:Genetic Complementation Test; D006801:Humans; D051379:Mice; D016513:Mice, SCID / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology; D014162:Transfection; D045567:VDJ Recombinases
D004254:DNA Nucleotidyltransferases; D045567:VDJ Recombinases