Analysis of 2-Chloro-2′-deoxyadenosine Incorporation into Cellular DNA by Quantitative Polymerase Chain Reaction

Yuh, Seon H.; Tibudan, Martin; Hentosh, Patricia

doi:10.1006/abio.1998.2782

Cited by 9 publications

(6 citation statements)

References 21 publications

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“…Previous reports have shown that the QPCR assay can be used to study cellular DNA damage in the specific genes after exposure to DNA damaging agents, such as cisplatin [33], carboplatin [33], nitrogen mustards [34,35], UV irradiation [36], chlorambucil [37], alkylbenzylguanine [38], and 2-chloro-2-deoxyadenosine [39]. QPCR is a reliable method, equivalently sensitive to the ICP-MS method to analyze the incorporation of platinum-based drugs into the gene of interest [33].…”

Section: Resultsmentioning

confidence: 99%

Altered DNA Binding and Amplification of Human Breast Cancer Suppressor Gene BRCA1 Induced by a Novel Antitumor Compound, [Ru(η6-p-phenylethacrynate)Cl2(pta)]

Chakree

Ovatlarnporn

Dyson

et al. 2012

IJMS

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The ruthenium-based complex [Ru(η6-p-phenylethacrynate)Cl2(pta)] (pta = 1,3,5-triaza-7-phosphatricyclo-[3.3.1.1]decane), termed ethaRAPTA, is an interesting antitumor compound. The elucidation of the molecular mechanism of drug activity is central to the drug development program. To this end, we have characterized the ethaRAPTA interaction with DNA, including probing the sequence specific modified DNA structural stability and DNA amplification using the breast cancer suppressor gene 1 (BRCA1) of human breast and colon adenocarcinoma cell lines as models. The preference of ethaRAPTA base binding is in the order A > G > T > C. Once modified, the ethaRAPTA-induced BRCA1 structure has higher thermal stability than the modified equivalents of its related compound, RAPTA-C. EthaRAPTA exhibits a higher efficiency than RAPTA-C in inhibiting BRCA1 amplification. With respect to both compounds, the inhibition of BRCA1 amplification is more effective in an isolated system than in cell lines. These data provide evidence that will help to understand the process of elucidating the pathways involved in the response induced by ethaRAPTA.

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Section: Resultsmentioning

confidence: 99%

Altered DNA Binding and Amplification of Human Breast Cancer Suppressor Gene BRCA1 Induced by a Novel Antitumor Compound, [Ru(η6-p-phenylethacrynate)Cl2(pta)]

Chakree

Ovatlarnporn

Dyson

et al. 2012

IJMS

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“…For example, T4 DNA polymerase does not incorporate diGTP opposite iC, whereas T7 RNA polymerase incorporates UTP but not iCTP opposite iG [15]. An isosteric analogue of iG, 2-chloroadenine, completely stops Taq polymerase when present in the template, which is rather unexpected, since many prokaryotic and eukaryotic polymerases accept the 2-chloroadenine-thymine pair, albeit to various extents [38].…”

Section: Figure 4 Tautomeric Forms Of Igmentioning

confidence: 99%

Neighbouring bases in template influence base-pairing of isoguanine

Maciejewska

Lichota

Kuśmierek

2003

Biochemical Journal

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Assuming that the efficiency of the incorporation of 5-methyl-2h-deoxyisocytosine-5h triphosphate (dMiCTP) and dTTP opposite isoguanine (iG) is a measure of the proportion of the keto and enol tautomers of iG in the Thermus aquaticus DNA polymerase active centre, we studied the influence of temperature and iGneighbouring bases in the template on base-pairing of iG. On the basis of experiments with four sequences (3h-TXT-5h, 3h-GXG-5h, 3h-CXC-5h, 3h-CXT-5h, where X l iG) at 37, 50, 65 and 80 mC, we found that 3h-neighbours decrease the fraction of the keto tautomer in the order C G T, whereas temperature apparently does not influence the tautomeric equilibrium of iG. The variability of the ratio of incorporation of dMiCTP versus dTTP (5-20) primarily reflects the variability of K m values, since

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“…The quantitative polymerase chain reaction (QPCR) was used to monitor the progress of Taq DNA polymerase utilizing DNA adducts as templates. Previous reports have shown that the QPCR method can be used to study cellular DNA damage and repair after exposure to DNA damaging agents such as 1, nitrogen mustards [41] [43 -45], 2-chloro-2-deoxyadenosine [46], chlorambucil [47], alkylbenzylguanine [48], and UV irradiation [44]. Such studies showed that the total amount of amplified PCR product from a given region is inversely proportional to the amount of DNA adducts within the specified DNA fragment, as long as the experimental conditions allow exponential amplification, and the concentration of the DNA template is not a limiting factor [44].…”

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confidence: 99%

In vitro Ruthenation of Human Breast Cancer Suppressor Gene 1 (BRCA1) by the Antimetastasis Compound RAPTA‐C and Its Analogue CarboRAPTA‐C

Ratanaphan

Temboot

Dyson

2010

Chemistry & Biodiversity

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The interaction of two ruthenium-arene-1,3,5-triaza-7-phosphaadamantane compounds ([Ru(eta(6)-p-cymene)Cl(2)(pta)] and [Ru(eta(6)-p-cymene)(C(6)H(6)O(4))(pta)], termed RAPTA-C (3) and carboRAPTA-C (4), resp.) with the DNA sequence of the human breast-cancer suppressor gene 1 (BRCA1) has been studied using a range of techniques that probe conformation, cross-linking, base specificity, restriction analysis, and in vitro inhibition of DNA polymerization. The study demonstrates that substitution of the two labile chloride ligands in 3 by the more stable cyclobutane-1,1-dicarboxylate ligand onto the RAPTA framework reduces the rate of reaction with DNA in a similar manner to the analogous Pt-based drug pair cisplatin (1) and carboplatin (2), suggesting that hydrolysis may be a prerequisite to DNA binding with the Ru compounds. Moreover, the rate of DNA interaction for 3 is in a similar range to that of 2, despite the fact that these compounds have a different therapeutic profile. The similar rates of reaction contrasting with the different modes of activity suggests that the RAPTA compounds may be clinically useful against cancer cells that have developed resistance to Pt-based therapies, particularly involving excision-repair mechanisms.

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Analysis of 2-Chloro-2′-deoxyadenosine Incorporation into Cellular DNA by Quantitative Polymerase Chain Reaction

Cited by 9 publications

References 21 publications

Altered DNA Binding and Amplification of Human Breast Cancer Suppressor Gene BRCA1 Induced by a Novel Antitumor Compound, [Ru(η6-p-phenylethacrynate)Cl2(pta)]

Altered DNA Binding and Amplification of Human Breast Cancer Suppressor Gene BRCA1 Induced by a Novel Antitumor Compound, [Ru(η6-p-phenylethacrynate)Cl2(pta)]

Neighbouring bases in template influence base-pairing of isoguanine

In vitro Ruthenation of Human Breast Cancer Suppressor Gene 1 (BRCA1) by the Antimetastasis Compound RAPTA‐C and Its Analogue CarboRAPTA‐C

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