A human gene that restores the DNA-repair defect in SCID mice is located on 8p11.1-->q11.1.
In order to map the gene that is responsible for the DNA-repair defect in severe combined immune deficient (SCID) mice, a mixture of microcells independently isolated from mouse A9 cells containing pSV2neo-tagged human chromosomes 5, 7, 8, 9, 11, 15, 18 or 20 were fused with SCID fibroblast cell lines SCVA2 and SCVA4, which were originally established from lung tissue of the C.B.17-scid/scid mouse by SV40 virus transfection. After irradiation with 60Co gamma-rays and selection with antibiotic G418, 12 independent clones were obtained, of which 4 contained an intact chromosome 8, 3 clones contained a deleted chromosome 8 [del(8)q22-->qter or del(8)q23--> qter] and remaining 5 had no detectable or specific human chromosome. We further independently transferred a single human chromosome 8 or 11 into the SCVA cells via microcell fusion, and examined the radiation sensitivity of the microcell hybrids. Complementation of the radiation sensitivity was correlated with the presence of human chromosome 8 in microcell hybrids, whereas no correlation was observed in clones following the transfer of human chromosome 11. Thus, the results indicate that human chromosome 8 restored high sensitivity to ionizing radiation. A number of subclones that were radiation resistant or sensitive were isolated from the microcell hybrids. The concordance of the radiation sensitivity with the presence or absence of specific DNA fragments on chromosome 8 indicates that the human gene is located on the centromeric region of chromosome 8, i.e., 8p11.1--> q11.1.
Kurimasa
A
A
Department of Molecular and Cell Genetics, School of Life Science, Faculty of Medicine, Tottori University, Japan.
Nagata
Y
Y
Shimizu
M
M
Emi
M
M
Nakamura
Y
Y
Oshimura
M
M
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002459:Cell Fusion; D002460:Cell Line; D002472:Cell Transformation, Viral / Q000528:radiation effects; D002478:Cells, Cultured; D002874:Chromosome Mapping; D002898:Chromosomes, Human, Pair 8; D003037:Cobalt Radioisotopes; D004247:DNA / Q000528:radiation effects; D004260:DNA Repair / Q000235:genetics* / Q000528:radiation effects; D005347:Fibroblasts; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011836:Radiation Tolerance / Q000235:genetics; D013539:Simian virus 40; D014162:Transfection
D003037:Cobalt Radioisotopes; D004247:DNA