Analysis of metal-induced mutations altering the expression or structure of a retroviral gene in a mammalian cell line.
Carcinogenic metal compounds, with the exception of chromium(VI), have been found to be poorly mutagenic in both prokaryotic and mammalian cell mutagenesis assays, yet they are clearly clastogenic (Hansen and Stern, 1984). Thus, the role of metals as initiators in carcinogenesis has been difficult to delineate. In an effort to develop a model system capable of assaying DNA damage caused by carcinogenic metals, we have investigated the role of NiCl2, CdCl2, Na2CrO4, and NMU in a murine sarcoma virus-infected mammalian cell line in which expression of the retroviral v-mos gene is growth-temperature regulated. This cell line, designated 6m2, contains a single-copy, stably integrated, mutant Moloney murine sarcoma virus DNA (designated MuSVts110) and is temperature sensitive for morphological transformation due to a conditionally defective viral RNA-splicing event that in turn regulates expression of the viral transforming gene. Mutations affecting the viral DNA in 6m2 cells can be detected if these alterations lead to changes in the structure or expression of the transforming protein encoded by the MuSVts110 v-mos gene. Analysis of the viral proteins from 6m2 'revertant' cell lines (as defined by reversion to the transformed phenotype at all growth temperatures) selected after treatment with the above agents showed that NiCl2, NMU, and Na2CrO4 each induced a different yet specific type of mutation. NiCl2 and NMU each altered the temperature sensitivity of viral RNA splicing, possibly due to base substitution mutations, but did so to distinctly different extents. Na2CrO4 affected the structure of the viral proteins by inducing what appear to be short frameshift mutations that resulted in the temperature-dependent translation of a novel virus-encoded transforming protein, P100gag-mos. CdCl2 also induced frameshift mutations but, in one case, induced a mutation which may result from a deletion of about 300 bases within the MuSVts110 DNA.
Biggart
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NW
Department of Tumor Biology, University of Texas M.D. Anderson Hospital and Tumor Institute, Houston 77030.
Murphy
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EC
D002460:Cell Line; D004279:DNA, Viral / Q000187:drug effects / Q000235:genetics; D015683:Gene Products, gag; D005796:Genes / Q000187:drug effects; D005814:Genes, Viral / Q000187:drug effects*; D008670:Metals / Q000494:pharmacology*; D008770:Methylnitrosourea / Q000494:pharmacology; D008980:Moloney murine sarcoma virus / Q000187:drug effects / Q000235:genetics*; D009154:Mutation; D017232:Oncogene Proteins v-mos; D009857:Oncogenes / Q000187:drug effects; D012191:Retroviridae Proteins / Q000235:genetics; D012492:Salts / Q000494:pharmacology; D009053:Sarcoma Viruses, Murine / Q000235:genetics*
D004279:DNA, Viral; D015683:Gene Products, gag; D008670:Metals; D017232:Oncogene Proteins v-mos; D012191:Retroviridae Proteins; D012492:Salts; D008770:Methylnitrosourea