AMP deaminase and thymidine kinase deficiencies in a mutant mouse S49 cell clone.
From a mutagenized population of wild type S49 cells, a clone was isolated in a single step that possessed functional and biochemical deficiencies in both AMP deaminase and thymidine kinase activities. This mutant cell line, DTB6, was selected in semi-solid medium containing 1mM thymidine and 1mM dibutyryl cyclic AMP. In comparative growth rate experiments, DTB6 cells were considerably less sensitive than parental cells to the growth inhibitory effects of thymidine. In contrast, DTB6 cells were much more sensitive to the cytotoxic effects of adenine and adenosine. The supersensitivity of DTB6 cells toward adenine could be ameliorated by the addition of hypoxanthine to the culture medium. The growth phenotype of the mutant cells could be attributed to deficiencies in two enzyme activities. First, DTB6 cells possessed a 60-70% deficiency in AMP deaminase activity, although the residual activity appeared kinetically similar to the wild type enzyme. Second, DTB6 cells possessed a virtual complete deficiency in thymidine kinase activity. Both enzyme deficiencies behaved in a recessive fashion in intraspecies hybrids. Revertants of DTB6 cells possessed wild type levels of AMP deaminase activity but remained deficient in thymidine kinase activity, while another revertant of DTB6 cells expressed 11% of the wild type thymidine kinase level but did not perceptibly change its AMP deaminase activity. The ability to isolate single step mutants with two seemingly independent biochemical abnormalities raises the speculation that there may be some link between cellular functions responsible for purine nucleotide and thymidine metabolism.
Hanson
S
S
Department of Biochemistry, Oregon Health Sciences University, Portland 97201.
Ullman
B
B
D000659:AMP Deaminase / Q000172:deficiency* / Q000235:genetics; D000225:Adenine / Q000633:toxicity; D000818:Animals; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002999:Clone Cells / Q000201:enzymology; D004305:Dose-Response Relationship, Drug; D006822:Hybrid Cells / Q000201:enzymology; D019271:Hypoxanthine; D007042:Hypoxanthines / Q000494:pharmacology; D008223:Lymphoma / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D009709:Nucleotide Deaminases / Q000172:deficiency*; D013936:Thymidine / Q000633:toxicity; D013937:Thymidine Kinase / Q000172:deficiency* / Q000235:genetics; D014407:Tumor Cells, Cultured
D007042:Hypoxanthines; D019271:Hypoxanthine; D013937:Thymidine Kinase; D009709:Nucleotide Deaminases; D000659:AMP Deaminase; D000225:Adenine; D013936:Thymidine