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Mutations that confer a hypersusceptibility to the hypoxanthine-guanine phosphoribosyltransferase-inhibiting compound N-(phosphonacetyl)-L-aspartate (PALA) in Chinese hamster ovary mutants.
A mutational approach was used to identify the gene encoding hypoxanthine-guanine phosphoribosyltransferase (HGPRT) in Chinese hamster ovary (CHO) cells. A cell line was constructed in which the guanine resistance gene was inactivated in the presence of increased amounts of guanine. This line, designated the G-RIB cell line, became highly sensitive to inhibition of HGPRT activity by N-(phosphonacetyl)-L-aspartate (PALA) and to the DNA-damaging effects of (1-3-C3H2PO2)2 [bis(p-chloro-m-xylene phosphonate)]. In the presence of PALA, the growth of the G-RIB cell line was inhibited dramatically. Thus, the G-RIB cell line has a genetically defined phenotype of spontaneous chromosomal aberrations and a hypersusceptibility to the cytotoxic effects of PALA. The spontaneous aberrations were suppressed in the G-RIB cell line by cultures in the presence of either increased amounts of guanine or chloramphenicol, suggesting that the G-RIB cells would be suitable for the rapid detection of PALA-induced mutations. After exposure of the cells to PALA, mutations were induced at a markedly higher frequency in G-RIB cells than in mutants containing the normal HGPR
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