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Purine reutilization and synthesis de novo in long-term human lymphocyte cell lines deficient in adenine phosphoribosyltransferase activity

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Somatic Cell Genetics

Abstract

Clonal lines, with either partial or total deficiency of adenine phosphoribosyltransferase (APRT) were derived from the WI-L2 long-term human lymphocyte line by selection for resistance to the adenine analogs 8-azaadenine or 2,6-diaminopurine. Resistance to 8-wazaadenine also conferred resistance to 2,6-diaminopurine and vice versa. Cells with 30–40% of wild-type APRT activity were selected by resistance to 0.01 mM 2,6-diaminopurine or 1.40 mM 8-azaadenine. The APRT in the 8-azaadinine-resistant cells exhibited a four- to sevenfold increase in the apparent Km for adenine. Activities of three other purine reutilization and interconversion enzymes in the resistant cells, including hypoxanthine phosphoribosyltransferase (HPRT), adenosine kinase, and adenosine deaminase, were within the range of wild-type activities. The doubling times of the APRT-deficient cells in purine-free medium was not different from wild-type cells. The APRT in the 8-azaadenine-resistant cells did not have an altered mobility in glycerol gradients as compared to wild-type cells. The rate of purine synthesis de novo and intracellular levels of 5-phosphoribosyl-1-pyrophosphate were unchanged in the APRT-deficient cells as compared to WI-L2. The ability of the cells to reutilize exogenous adenine, however, was severely impaired.

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Author notes

  1. Elaine B. Spector

    Present address: Mental Retardation Research Center, Department of Psychiatry, University of California Los Angeles Medical School, 90024, Los Angeles, California

  2. Michael S. Hershfield

    Present address: Department of Medicine, Duke University School of Medicine, 27710, Durham, North Carolina

Authors and Affiliations

  1. Department of Medicine, University of California San Diego Medical School, 92093, La Jolla, California

    Elaine B. Spector, Michael S. Hershfield & J. Edwin Seegmiller

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  1. Elaine B. Spector
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  2. Michael S. Hershfield
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  3. J. Edwin Seegmiller
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Spector, E.B., Hershfield, M.S. & Seegmiller, J.E. Purine reutilization and synthesis de novo in long-term human lymphocyte cell lines deficient in adenine phosphoribosyltransferase activity. Somat Cell Mol Genet 4, 253–264 (1978). https://doi.org/10.1007/BF01542842

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  • DOI: https://doi.org/10.1007/BF01542842

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