Literature DB >> 20005183

Use of Drosophila deoxynucleoside kinase to study mechanism of toxicity and mutagenicity of deoxycytidine analogs in Escherichia coli.

Brittany Betham1, Sophia Shalhout, Victor E Marquez, Ashok S Bhagwat.   

Abstract

Most bacteria, including Escherichia coli, lack an enzyme that can phosphorylate deoxycytidine and its analogs. Consequently, most studies of toxicity and mutagenicity of cytosine analogs use ribonucleosides such as 5-azacytidine (AzaC) and zebularine (Zeb) instead of their deoxynucleoside forms, 5-aza-2'-deoxycytidine (AzadC) and 2'-deoxy-zebularine (dZeb). The former analogs are incorporated into both RNA and DNA creating complex physiological responses in cells. To circumvent this problem, we introduced into E. coli the Drosophila deoxynucleoside kinase (Dm-dNK), which has a relaxed substrate specificity, and tested these cells for sensitivity to AzadC and dZeb. We find that Dm-dNK expression increases substantially sensitivity of cells to these analogs and dZeb is very mutagenic in cells expressing the kinase. Furthermore, toxicity of dZeb in these cells requires DNA mismatch correction system suggesting a mechanism for its toxicity and mutagenicity. The fluorescence properties of dZeb were used to quantify the amount of this analog incorporated into cellular DNA of mismatch repair-deficient cells expressing Dm-dNK and the results showed that in a mismatch correction-defective strain a high percentage of DNA bases may be replaced with the analog without long term toxic effects. This study demonstrates that the mechanism by which Zeb and dZeb cause cell death is fundamentally different than the mechanism of toxicity of AzaC and AzadC. It also opens up a new way to study the mechanism of action of deoxycytidine analogs that are used in anticancer chemotherapy. Copyright 2009 Elsevier B.V. All rights reserved.

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Year:  2009        PMID: 20005183      PMCID: PMC2819640          DOI: 10.1016/j.dnarep.2009.11.006

Source DB:  PubMed          Journal:  DNA Repair (Amst)        ISSN: 1568-7856


  38 in total

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Journal:  Mutat Res       Date:  1986-12       Impact factor: 2.433

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Journal:  Biochim Biophys Acta       Date:  1973-09-28

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Journal:  Mutat Res       Date:  1983-07       Impact factor: 2.433

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Journal:  Proc Natl Acad Sci U S A       Date:  1980-02       Impact factor: 11.205

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Journal:  Mol Gen Genet       Date:  1978-07-25

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Journal:  Proc Natl Acad Sci U S A       Date:  1986-02       Impact factor: 11.205

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Authors:  J Neuhard
Journal:  J Bacteriol       Date:  1968-11       Impact factor: 3.490

10.  Potent inhibition of HhaI DNA methylase by the aglycon of 2-(1H)-pyrimidinone riboside (zebularine) at the GCGC recognition domain.

Authors:  Victor E Marquez; Ramon Eritja; James A Kelley; Dana Vanbemmel; Judith K Christman
Journal:  Ann N Y Acad Sci       Date:  2003-12       Impact factor: 5.691

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  2 in total

1.  Exploring Synergy between Classic Mutagens and Antibiotics To Examine Mechanisms of Synergy and Antibiotic Action.

Authors:  Lisa Yun Song; Sara D'Souza; Karen Lam; Tina Manzhu Kang; Pamela Yeh; Jeffrey H Miller
Journal:  Antimicrob Agents Chemother       Date:  2015-12-28       Impact factor: 5.191

2.  Repair of DNA Damage Induced by the Cytidine Analog Zebularine Requires ATR and ATM in Arabidopsis.

Authors:  Chun-Hsin Liu; Andreas Finke; Mariana Díaz; Wilfried Rozhon; Brigitte Poppenberger; Tuncay Baubec; Ales Pecinka
Journal:  Plant Cell       Date:  2015-05-28       Impact factor: 11.277

  2 in total

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