Based on recent progress in our ability to amplify mutant DNA sequences by high fidelity polymerase chain reaction and to separate mutant and wild type DNA sequences using denaturing gradient electrophoresis, we propose three separate efforts, each necessary to the intended use of this new technology to study the rate of combustion effluents in mutation in humans. First, we need to understand the relationship between existing cellular modes of analysis of mutation in human cell samples and our proposed molecular mode. At present, mutants are enumerated as 6-thioguanine resistant cells, growing as colonies. These mutants have little or not activity for the enzyme hypoxanthine phosphoribosyl transferase (hpt). We have developed an approach which examines point mutations at the DNA sequence level in one or a few exons of hprt. These independent modes reveal partially overlapping sets of mutations, and their relationship requires careful definition. We propose to use the mutagens fluoranthene, cyclopenta(c,d)pyrene and benzo(alpha)pyrene in this characterization effort. Although not a major mutagen in incomplete combustion effluents, benzo(alpha)pyrene's mutational spectrum has been studied in bacteria, and our effort will permit an important human cell/bacterial comparison. Secondly and thirdly, we are combining with Projects C-1, C-2 and D-3 to study the quantitative dose and time dependence of mutation in human cells and mutation and tumor formation in mouse tissue. These studies will result in understanding of the relationships in time among protein adducts, DNA adducts and mutation after exposures to fluoranthene, cyclopenta(c,d)pyrene, benzo(alpha)pyrene and important pyrolysis products as shall be determined by research in Projects A-1,-2,-3,-4, B and D-1,-2, and -3.

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National Institute of Environmental Health Sciences (NIEHS)
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Massachusetts Institute of Technology
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Tomita-Mitchell, Aoy; Ling, Losee Lucy; Glover, Curtis L et al. (2003) The mutational spectrum of the HPRT gene from human T cells in vivo shares a significant concordant set of hot spots with MNNG-treated human cells. Cancer Res 63:5793-8
Tomita-Mitchell, A; Kat, A G; Marcelino, L A et al. (2000) Mismatch repair deficient human cells: spontaneous and MNNG-induced mutational spectra in the HPRT gene. Mutat Res 450:125-38
Durant, J L; Lafleur, A L; Busby Jr, W F et al. (1999) Mutagenicity of C24H14 PAH in human cells expressing CYP1A1. Mutat Res 446:1-14
Wang, J S; He, X; Mulder, P P et al. (1999) Comparative tumorigenicity of the cyclopenta-fused polycyclic aromatic hydrocarbons aceanthrylene, dihydroaceanthrylene and acephenanthrylene in preweanling CD-1 and BLU:Ha mouse bioassays. Carcinogenesis 20:1137-41
Busby Jr, W F; Smith, H; Crespi, C L et al. (1997) Mutagenicity of the atmospheric transformation products 2-nitrofluoranthene and 2-nitrodibenzopyranone in Salmonella and human cell forward mutation assays. Mutat Res 389:261-70
Busby Jr, W F; Smith, H; Plummer, E F et al. (1997) Mutagenicity of cyclopenta-fused polynuclear aromatic hydrocarbons and a non-polar fraction from a fuel combustion sample in a Salmonella forward mutation assay without exogenous metabolic activation. Mutat Res 391:117-25
Durant, J L; Busby Jr, W F; Lafleur, A L et al. (1996) Human cell mutagenicity of oxygenated, nitrated and unsubstituted polycyclic aromatic hydrocarbons associated with urban aerosols. Mutat Res 371:123-57
Palotas, A B; Rainey, L C; Feldermann, C J et al. (1996) Soot morphology: an application of image analysis in high-resolution transmission electron microscopy. Microsc Res Tech 33:266-78
Wang, J S; Busby Jr, W F (1996) Bacterial and human cell mutagenicity and mouse lung tumorigenicity of the oxygenated polynuclear aromatic hydrocarbon phenalenone. Fundam Appl Toxicol 33:212-9
Wang, J S; Busby, W F; Wogan, G N (1995) Tissue distribution of DNA adducts in pre-weanling BLU:Ha mice treated with a tumorigenic dose of fluoranthene. Cancer Lett 92:9-19

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