Mutagenesis induced by site specifically placed 4'-hydroxymethyl-4,5',8-trimethylpsoralen adducts

Piette, Jacques; Gamper, Howard; Vorst, A. Van de; Hearst, John E.

doi:10.1093/nar/16.21.9961

Cited by 29 publications

(28 citation statements)

References 44 publications

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“…The predicted nature of the mutations was confirmed by sequencing the plasmids extracted from clones chosen on the basis of their hybridization profile. Among 54 mutant plasmids sequenced, all were found to be targeted to the site of the adduct, thus confirming an observation made in other single-adduct mutagenesis studies (3,6,40). More precisely, all mutant plasmids had a base substitution involving the adenine residue of the cisDDP{d(ApG)} adduct, as expected from previous mutation spectrum studies (23).…”

supporting

confidence: 83%

Single d(ApG)/cis-diamminedichloroplatinum(II) adduct-induced mutagenesis in Escherichia coli.

Burnouf

Gauthier

Chottard

et al. 1990

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

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The mutation spectrum induced by the widely used antitumor drug cis-diamminedichloroplatinum(II) (cis-DDP) showed that cisDDP{d(ApG)} adducts, although they account for only 25% of the lesions formed, are =5 times more mutagenic than the major GG adduct. We report the construction of vectors bearing a single cisDDP{d(ApG)} lesion and their use in mutagenesis experiments in Escherichia coli. The mutagenic processing of the lesion is found to depend strictly on induction of the SOS system of the bacterial host cells. In SOS-induced cells, mutation frequencies of 1-2% were detected. All these mutations are targeted to the 5' base of the adduct. Single A -+ T transversions are mainly observed (80%), whereas A -* G transitions account for 10% of the total mutations. Tandem base-pair substitutions involving the adenine residue and the thymine residue immediately 5' to the adduct occur at a comparable frequency (10%). No selective loss of the strand bearing the platinum adduct was seen, suggesting that, in vivo, cisDDP{d(ApG)} adducts are not blocking lesions. The high mutation specificity of cisDDP{d(ApG)}-induced mutagenesis is discussed in relation to structural data.During the past decade, progress in molecular biology allowed the development of strategies for elucidating the molecular mechanisms involved in processing the damages introduced in DNA by physical or chemical agents. A general procedure is to determine the forward mutation spectrum induced by a particular agent on a target DNA sequence (1). This approach allows definition of the general characteristics of the mutagenic processes, such as their genetic requirements and their specificity. However, these strategies are not sensitive enough to measure the relative contribution of different adducts in repair or mutagenesis events. This problem has been overcome by the use of synthetic oligodeoxyribonucleotides containing defined adducts located at precise sites (for review, see ref.2). When these adducts are introduced in a replicative vector and transfected into cells, several biological endpoints can be monitored: toxicity (3, 4), mutation efficiency and specificity (3-6), effect of the repair genotype of the host (7), and effect of surrounding sequences (3, 6). Furthermore, such modified oligonucleotides became useful tools for studying the structure of the DNA helix in the vicinity of the adduct (8-13) and precise biochemistry of repair (14-16).cis-Diamminedichloroplatinum(II) (cisDDP) is a widely used antitumor drug that binds to DNA, its main cellular target, by preferentially forming intrastrand crosslinks between the N7 atoms of adjacent purines (17)(18)(19). At low modification levels, the main adducts result from the chelation of two adjacent guanines [cisDDP{d(GpG)}], 65% of total lesions), whereas 25% of the adducts arise at 5' d(ApG) sequences. d(GpHpG) adduct (H = A, C, or T) and interstrand crosslinks altogether represent <10% of the total lesions (20-22).We have described the spectrum of mutations induced by cisDDP in Escherichia c...

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supporting

confidence: 83%

Single d(ApG)/cis-diamminedichloroplatinum(II) adduct-induced mutagenesis in Escherichia coli.

Burnouf

Gauthier

Chottard

et al. 1990

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

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“…TA + GC transversions are also recovered. 133 In view of more recent data,75. '34 a transient misalignment model might better explain the base substitutions as well as the frameshift events observed.…”

Section: Furocoumarin-induced Mutationmentioning

confidence: 99%

Distribution and Repair of Photolesions in Dna: Genetic Consequences and the Role of Sequence Context

Sage

1993

Photochem & Photobiology

226

108

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“…Denaturing electrophoresis and autoradiography (autoradiograph in Fig. 4A, fractions [21][22][23][24][25][26][27][28][29][30][31][32][33][34] revealed the presence of predominantly three DNA species. The first peak area (fractions [21][22][23][24][25][26] contained mostly 8UM and very little of 8XL and 8-mer pyrone-side monoadduct (8Mpy).…”

Section: Resultsmentioning

confidence: 99%

“…The first peak area (fractions [21][22][23][24][25][26] contained mostly 8UM and very little of 8XL and 8-mer pyrone-side monoadduct (8Mpy). The second peak area (fractions [29][30][31][32][33][34] represented mostly 8XL and relatively smaller amounts of Mpy and 8UM. In addition to 8XL some of these fractions contained minute amounts of DNA molecules migrating much slower than 8XL on these gels (data not shown).…”

Section: Resultsmentioning

confidence: 99%

“…We were able to synthesize 3 jumol of >95% pure 8XL (Fig. 5 (27,28), elongation complexes of RNA polymerases (12)(13)(14)(15), screening of complex genomes (29), induced mutagenesis (30), and DNA structure and dynamics (8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18). There is a clear need to solve the molecular structure of monoadducted and cross-linked DNA molecules by NMR or x-ray crystallography.…”

Section: Resultsmentioning

confidence: 99%

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Methods for the large-scale synthesis of psoralen furan-side monoadducts and diadducts.

Spielmann

Sastry

Hearst

1992

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

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We report methods for the preparation of multimicromole quantities of extremely pure uniquely photoadducted psoralen-DNA furan-side monoadducts and diadducts (cross-links). The methods use high-intensity krypton and argon ion lasers in the photoreactions and HPLC methods to purify the required oligonucleotides containing the photoadducts. With these methods we have syntheszed 2-3 #&mol of 8-mer psoralen furan-side monoadduct and diadduct. These methods allow one to generate large amounts of psoralenated DNA oligonucleotides and facilitate their study by NMR and x-ray crystallography.Psoralens are linear furocoumarins that have been used for the treatment of certain types of skin diseases (1, 2), antileukemia therapy (photophoresis), and the sterilization of blood products (3, 4). Psoralens photochemically alkylate DNA by reacting with pyrimidines (5). Fig. 1 shows the structure of the three main adducts between thymidine and HMT (5, 6). Reactions take place at the 3,4 or 4',5' double bonds of the psoralen with the 5,6 double bond in pyrimidines. The first step is the intercalation of the planar psoralen between base pairs of a double-stranded nucleic acid. Psoralen then reacts with the pyrimidine bases to form a MAf or a Mpy when UV-irradiated at a long wavelength (320-410 nm). By absorbing a second photon, the MAf can photoreact with adjacent pyrimidine bases on the opposite strand of doublestranded DNA (or RNA) to form an interstrand XL. Mpys do not absorb light at wavelengths above 320 nm and cannot be driven to form interstrand XLs with long-wavelength UV irradiation. All the photochemical adducts can be photoreversed by exposure to 254-nm UV or, for MAfs and XLs, by treatment with base (7,8). A solution structure derived from NMR data for the 8-mer 5'-d(GGGTACCC)-3' cross-linked with the psoralen 4'-aminomethyl-4,5',8-trimethylpsoralen has been reported (9).Psoralens have been very useful as probes for nucleic acid structure and function (10,11). Transcriptional elongation complexes arrested by psoralen adducts at specific sites on a DNA template have been generated (12-15). Psoralen-DNA adducts are substrates for DNA repair enzymes (16). Other types of protein-DNA interactions have also been studied using psoralens as probes (17-21).The lack of methods for the large-scale synthesis of psoralen monoadducts and diadducts is a major unsolved problem in the research on psoralenated DNA oligonucleotides. Traditional methods used Hg/Xe arc lamps (22) and blackray light (23) for investigation of the photochemistry of psoralen-DNA interactions. Photochemical efficiency with these light sources is very low and often results in the generation of multiple photoproducts, despite the use of appropriate filters to achieve monochromicity. Traditional methods rely on denaturing PAGE followed by DNA elution from gel slices to separate the photoadducted DNA oligonucleotides from other components in the reaction mixture. This procedure results in considerable loss of DNA during sample work-up and the persistence of c...

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Mutagenesis induced by site specifically placed 4'-hydroxymethyl-4,5',8-trimethylpsoralen adducts

Cited by 29 publications

References 44 publications

Single d(ApG)/cis-diamminedichloroplatinum(II) adduct-induced mutagenesis in Escherichia coli.

Single d(ApG)/cis-diamminedichloroplatinum(II) adduct-induced mutagenesis in Escherichia coli.

Distribution and Repair of Photolesions in Dna: Genetic Consequences and the Role of Sequence Context

Methods for the large-scale synthesis of psoralen furan-side monoadducts and diadducts.

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