Complementation of a DNA repair defect in xeroderma pigmentosum cells by transfer of human chromosome 9.
Complementation of the repair defect in xeroderma pigmentosum cells of complementation group A was achieved by the transfer of human chromosome 9. A set of mouse-human hybrid cell lines, each containing a single Ecogpt-marked human chromosome, was used as a source of donor chromosomes. Chromosome transfer to XPTG-1 cells, a hypoxanthine/guanine phosphoribosyltransferase-deficient mutant of simian virus 40-transformed complementation group A cells, was achieved by microcell fusion and selection for Ecogpt. Chromosome-transfer clones of XPTG-1 cells, each containing a different human donor chromosome, were analyzed for complementation of sensitivity to UV irradiation. Among all the clones, increased levels of resistance to UV was observed only in clones containing chromosome 9. Since our recipient cell line XPTG-1 is hypoxanthine/guanine phosphoribosyltransferase deficient, cultivation of Ecogpt+ clones in medium containing 6-thioguanine permits selection of cells for loss of the marker and, by inference, transferred chromosome 9. Clones isolated for growth in 6-thioguanine, which have lost the Ecogpt-marked chromosome, exhibited a UV-sensitive phenotype, confirming the presence of the repair gene(s) for complementation group A on chromosome 9.
Kaur
G P
GP
Department of Microbiology and Molecular Genetics, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, Newark 07103-2757.
Athwal
R S
RS
D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000528:radiation effects; D002899:Chromosomes, Human, Pair 9; D002999:Clone Cells; D004260:DNA Repair; D004261:DNA Replication / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D005816:Genetic Complementation Test; D006801:Humans; D006822:Hybrid Cells / Q000166:cytology; D007041:Hypoxanthine Phosphoribosyltransferase / Q000172:deficiency / Q000235:genetics; D007621:Karyotyping; D008677:Metaphase; D051379:Mice; D014162:Transfection; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum / Q000235:genetics*
D007041:Hypoxanthine Phosphoribosyltransferase