Optically active benzo[c]phenanthrene diol epoxides bind extensively to adenine in DNA

Dipple, Anthony; Pigott, Margaret A.; Agarwal, Shiv Kumar; Yagi, Haruhiko; Sayer, Jane M.; Jerina, Donald M.

doi:10.1038/327535a0

Cited by 148 publications

(185 citation statements)

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“…4). Similarly, BcPHDE is an aromatic hydrocarbon that can form adducts with A or G (7,22). The action of this mutagen has been studied at the dhfr locus in CHO cells; both A and G were targets, with a great preference for the 5' base of di-or tripurine sequences (13).…”

Section: Resultsmentioning

confidence: 99%

Direct selection for mutations affecting specific splice sites in a hamster dihydrofolate reductase minigene.

Chen¹,

Chasin²

1993

Mol. Cell. Biol.

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A Chinese hamster cell line containing an extra exon 2 (50 bp) inserted into a single intron of a dihydrofolate reductase (dhfr) minigene was constructed. The extra exon 2 was efficiently spliced into the RNA, resulting in an mRNA that is incapable of coding for the DHFR enzyme. Mutations that decreased splicing of this extra exon 2 caused it to be skipped and so produced normal dhfr mRNA. In contrast to the parental cell line, the splicing mutants display a DHFR-positive growth phenotype. Splicing mutants were isolated from this cell line after treatment with four different mutagens (racemic benzo[clphenanthrene diol epoxide, ethyl methanesulfonate, ethyl nitrosourea, and UV irradiation). By polymerase chain reaction amplification and direct DNA sequencing, we determined the base changes in 66 mutants. Each of the mutagens generated highly specific base changes. All mutations were single-base substitutions and comprised 24 different changes distributed over 16 positions. Most of the mutations were within the consensus sequences at the exon 2 splice donor, acceptor, and branch sites. The RNA splicing patterns in the mutants were analyzed by quantitative reverse transcriptionpolymerase chain reaction. The recruitment of cryptic sites was rarely seen; simple exon skipping was the predominant mutant phenotype. The wide variety of mutations that produced exon skipping suggests that this phenotype is the typical consequence of splice site damage and supports the exon definition model of splice site selection. A few mutations were located outside the consensus sequences, in the exon or between the branch point and the polypyrimidine tract, identifying additional positions that play a role in splice site definition. That most of these 66 mutations fell within consensus sequences in this near-saturation mutagenesis suggests that splicing signals beyond the consensus may consist of robust RNA structures.The development of cell-free splicing systems has led to rapid and continuing progress in understanding biochemical mechanisms involved in intron removal and in the definition of the spliceosomal machinery. However, the exact requirements for recognition of splices sites and the rules governing splice site selection remain unclear. Analysis of consensus sequences provided the first definition of splice sites (49). Most efforts to move beyond that initial step have focused on site-directed mutagenesis studies (for reviews, see references 4 and 31). In these experiments, the effects of splice site mutations are typically assayed by using cell-free splicing extracts (in. vitro) or after transient transfection of cultured mammalian cells or injection of RNA directly into frog oocytes (in vivo).Conflicting results between these in vitro and in vivo assays for the effect of mutations on splicing have frequently been reported (2,38,39,45,83), raising the possibility that artifacts are being introduced in one system or the other. Moreover, recent experiments suggesting that the splicing machinery may be compartmentalized within t...

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Section: Resultsmentioning

confidence: 99%

Direct selection for mutations affecting specific splice sites in a hamster dihydrofolate reductase minigene.

Chen¹,

Chasin²

1993

Mol. Cell. Biol.

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“…For example, (-)-BcPhDE-1 is essentially devoid of tumorigenic activity, whereas (-)-BcPhDE-2 is the most active hydrocarbon dihydrodiol epoxide tested to date, and the other stereoisomers exhibit intermediate activities (12). To investigate the basis for these tumorigenic activities, relative mutagenicities in bacterial and mammalian cells have been reported (13), as have detailed studies of the chemistry of their interactions with DNA (14,15). These studies have shown that the exocyclic amino groups of deoxyadenosine and deoxyguanosine are the principal sites of reaction and that opening of the dihydrodiol epoxide occurs both cis and trans at the benzylic position.…”

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confidence: 99%

Mutagenic specificities of four stereoisomeric benzo[c]phenanthrene dihydrodiol epoxides.

Bigger

John

Yagi

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

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“…Other planar PAH diol-epoxides that contain a bay or fjord region, such as benz [a]anthracene and chrysene, bind covalently and almost exclusively with guanine bases in DNA. By contrast, the fjord region diol-epoxides of benzo[c]phenanthrene, with a distortion from the planarity of the parent molecule, react with adenine and guanine bases in DNA (Dipple et al, 1987;Jerina et al, 1991). The poorly repaired DNA adducts can persist until replication and cause mutations (Smith et al, 2000;Feng et al, 2002;Rodríguez et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Evaluation of the noncovalent binding interactions between polycyclic aromatic hydrocarbon metabolites and human p53 cDNA

Wei¹,

Lin²,

Zhang³

et al. 2010

Science of The Total Environment

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The binding of reactive polycyclic aromatic hydrocarbon (PAH) metabolites, formed enzymatically, to DNA is a crucial step in PAH carcinogenesis in vivo. We investigated the noncovalent binding interactions between 11 PAH metabolites and human p53 complementary DNA (p53 cDNA) using the fluorescence displacement method and molecular docking analysis. All of the examined metabolites predominantly interacted with p53 cDNA by intercalation instead of groove binding. The dissociation constants ranged from 0.02 to 12.34 μM. Of the metabolites tested, 1-hydroxypyrene and 3-hydroxybenzo[a]pyrene showed the strongest binding affinities to DNA, while 2-naphthol was the weakest DNA intercalator. The intercalation of the metabolites was stabilized by stacking the PAH phenyl rings with the DNA base pairs and the formation of hydrogen bonds between the oxide or hydroxyl groups on the metabolites, and DNA bases or backbones. The binding of the metabolites to DNA showed some sequence selectivity. The binding affinities and hydrogen bonds for 3-hydroxybenzo[a]pyrene, benzo[a]pyrene-4,5-dihydroepoxide (BPE) and benzo[a]pyrene-r-7,t-8-dihydrodiol-t-9,10-epoxide (BPDE) differed. It seems that the functional groups on the periphery of the PAH aromatic ring play crucial roles in regulating its binding affinity with DNA. Although it was difficult to determine the correlation between DNA noncovalent binding affinity and carcinogenicity for some of the PAH metabolites, the present study improved our understanding of the formation of PAH metabolite-DNA adducts.

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Optically active benzo[c]phenanthrene diol epoxides bind extensively to adenine in DNA

Cited by 148 publications

References 0 publications

Direct selection for mutations affecting specific splice sites in a hamster dihydrofolate reductase minigene.

Direct selection for mutations affecting specific splice sites in a hamster dihydrofolate reductase minigene.

Mutagenic specificities of four stereoisomeric benzo[c]phenanthrene dihydrodiol epoxides.

Evaluation of the noncovalent binding interactions between polycyclic aromatic hydrocarbon metabolites and human p53 cDNA

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