A gene correcting the defect in the CHO mutant Ade -H, deficient in a branch point enzyme (adenylosuccinate synthetase) of de novo purine biosynthesis, is located on the long arm of chromosome 1.
Somatic hybrids between human cells and the Chinese hamster ovary (CHO) K1 mutant, Ade -H cells, were selected for purine prototrophy by growth in adenine-free medium. The Ade -H mutant is defective in the enzyme adenylosuccinate (AMPS) synthetase (ADSS; EC 6.3.4.4), which carries out the first of a two-step sequence in the biosynthesis of AMP from IMP, and therefore requires exogenous adenine for growth. The presence of the long arm of human chromosome 1 in the hybrids is 100% concordant for the ability to grow in adenine-free medium and restoration of the enzyme activity. Hybrid segregants that lose the ability to grow in adenine-free medium lose all or a portion of chromosome 1 and enzyme activity. Southern blot hybridization with a chromosome 1-specific probe, BCMI, confirms the existence of human chromosome 1 in these hybrids. Analysis of a human/CHO translocation chromosome that arose in one of the hybrids suggests that the gene correcting the defect lies in the region 1 cen-1q12. In summary, we have shown by cytogenetics, segregant analysis, biochemical assay, and Southern blot analysis that human chromosome 1, most likely in the region 1cen-1q12, corrects the defect in ADSS-deficient mutant Ade-H cells.
Lai
L W
LW
Eleanor Roosevelt Institute for Cancer Research, Denver, Colorado 80206.
Hart
I M
IM
Patterson
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D000265:Adenylosuccinate Synthase / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D015139:Blotting, Southern; D002460:Cell Line; D002878:Chromosomes, Human, Pair 1; D006224:Cricetinae; D004247:DNA / Q000032:analysis; D006801:Humans; D006822:Hybrid Cells; D009154:Mutation; D011687:Purines / Q000378:metabolism*
D011687:Purines; D004247:DNA; D000265:Adenylosuccinate Synthase