Synthesis of 5-chlorodeoxyuridine and a comparative study of 5-halodeoxyuridines in E. Coli

Visser, Donald W.; Frisch, David M.; Huang, Bing

doi:10.1016/0006-2952(60)90017-4

Cited by 38 publications

(16 citation statements)

References 10 publications

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“…The damaging agents include hydrogen peroxide and hypochlorous acid resulting in both oxidized and chlorinated bases. Among these damage products is 5-chlorouracil (ClU), which could arise by chlorination and deamination of cytosine residues in DNA and nucleotides or chlorination of uracil bases present in nucleotide precursor pools (6–10) as shown in Fig. 1.…”

Section: Introductionmentioning

confidence: 99%

Base Pairing Configuration and Stability of an Oligonucleotide Duplex Containing a 5-Chlorouracil-Adenine Base Pair

et al. 2009

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Inflammation-mediated reactive molecules can damage DNA by oxidation and chlorination. The biological consequences of this damage are as yet incompletely understood. In this paper, we have constructed oligonucleotides containing 5-chlorouracil (ClU), one of the known inflammation damage products. The thermodynamic stability, base pairing configuration and duplex conformation of oligonucleotides containing ClU paired opposite adenine have been examined. NMR spectra reveal that the ClU-A base pair adopts a geometry similar to that of the T-A base pair, and the ClU-A containing duplex adopts a normal B-form conformation. The linewidth of the imino proton of the ClU residue is substantially greater than that of the corresponding T imino proton; however, this difference is not attributed to a reduced thermal or thermodynamic stability or to increased proton exchange with solvent. While the NMR studies reveal increased chemical exchange for the ClU imino proton of the ClU-A base pair, the ClU residue is not a target for removal by the Escherichia coli mispaired uracil glycosylase, which senses damage-related helix instability. The results of this study are consistent with previous reports indicating that the DNA of replicating cells can tolerate substantial substitution with CIU. The fraudulent, pseudo-Watson-Crick ClU-A base pair is sufficiently stable to avoid glycosylase removal and, therefore, might constitute a persistent form of cellular DNA damage.

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Section: Introductionmentioning

confidence: 99%

Base Pairing Configuration and Stability of an Oligonucleotide Duplex Containing a 5-Chlorouracil-Adenine Base Pair

et al. 2009

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“…[14] We chose chlorouracil (c) as the most promising candidate for DNAbase transliteration (see Figure S1b) because: 1) it closely resembles thymine in structural studies of synthetic DNA duplexes, which suggests that the stability of the A:c pair is close to that of A:T; [15] 2) it is readily converted into the chlorodeoxyuridine nucleoside and the dcMP, dcDP, and dcTP nucleotides by nucleoside phosphorylases, thymidine kinase, thymidylate kinase, and nucleoside diphosphate kinase, respectively; [2] 3) unlike fluorouracil, it is less liable to elevated tautomerism, which causes ambiguous pairing with guanine as well as adenine; [15] 4) unlike the bromo and iodo analogues, it is not reduced to uracil at the redox potential of the cytoplasm. [16] Chlorine is present in numerous natural compounds, especially secondary metabolites of marine organisms, and a number of enzymatic mechanisms that enable the formation of C À Cl bonds have been elucidated. [17] Apparently, it has not been reported to occur in any natural nucleic acid building block so far.…”

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confidence: 99%

“…The effects for both analogues have been described as very similar and encompass filamentation, cessation of growth, low mortality, and mutagenesis. [16,18] The strain THY1 that was subjected to 5-chlorouracil transliteration is a derivative of wild-type E. coli K12 strain MG1655. [19] Its construction involved consecutive deletions of the genes thyA and udp and the deoCABD operon from the MG1655 chromosome, and the insertion of a P15A plasmid carrying the Lactobacillus leichmannii gene ntd, which encodes nucleoside deoxyribosyltransferase (see Supporting Information).…”

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confidence: 99%

Chemical Evolution of a Bacterium’s Genome

Marlière¹,

et al. 2011

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Automated selection was used to evolve an Escherichia coli strain unable to synthesize thymine nucleotides into a chemically modified organism whose DNA genome is composed of adenine, cytosine, guanine, and an artificial base, the thymine analogue 5‐chlorouracil. Evolving cells were initially observed as irregular filaments and progressively recovered the appearance of short rods typical of wild‐type E. coli (see picture).

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“…We observe that 10 µM concentration of CldU in the media does not alter cell division kinetics; however, the mole ratio of ClU to T in the DNA is 0.89 ± 0.02 when cells are grown for two cell doublings. Previously it has been shown that CldU is metabolized and incorporated into DNA using antibodies that bind selectively to DNA containing halogenated bases (28–32). We have confirmed and extended these finding using a mass spectral method as reported here.…”

Section: Discussionmentioning

confidence: 99%

“…While it is known that 5-chloro-2’-deoxyuridine (CldU) added to culture media is metabolized and incorporated into DNA as shown by antibody binding (28–32), CldU induces senescence, toxicity, and sister-chromatid exchanges by as yet unknown mechanisms (33–36). …”

Section: Introductionmentioning

confidence: 99%

Polymerase Incorporation and Miscoding Properties of 5-Chlorouracil

Kim

Darwanto

Theruvathu

et al. 2010

Chem. Res. Toxicol.

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Inflammation-mediated hypochlorous acid (HOCl) can damage DNA, DNA precursors, and other biological molecules, thereby producing an array of damage products such as 5-chlorouracil (ClU). In this study, we prepared and studied 5-chloro-2’-deoxyuridine (CldU) and ClU-containing oligonucleotide templates. We demonstrate that human K-562 cells grown in culture with 10 µM CldU incorporate substantial amounts of CldU without significant toxicity. When in the template, ClU residues pair with dATP, but also with dGTP, in a pH-dependent manner with incorporation by human polymerase β, Avian Myeloblastosis Virus reverse transcriptase (AMV-RT), and Escherichia coli Klenow fragment (exo−) polymerase. The enhanced miscoding of ClU is attributed to the electron-withdrawing 5-chlorine substituent that promotes formation of an ionized ClU-G mispair. When mispaired with G, ClU is targeted for removal by human glycosylases. The formation, incorporation and repair of ClU could promote transition mutations and other forms of heritable DNA damage.

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Synthesis of 5-chlorodeoxyuridine and a comparative study of 5-halodeoxyuridines in E. Coli

Cited by 38 publications

References 10 publications

Base Pairing Configuration and Stability of an Oligonucleotide Duplex Containing a 5-Chlorouracil-Adenine Base Pair

Base Pairing Configuration and Stability of an Oligonucleotide Duplex Containing a 5-Chlorouracil-Adenine Base Pair

Chemical Evolution of a Bacterium’s Genome

Polymerase Incorporation and Miscoding Properties of 5-Chlorouracil

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