Enhanced Cytotoxicity of Nucleoside Analogs by Overexpression of Mitochondrial Deoxyguanosine Kinase in Cancer Cell Lines

Zhu, Changliang; Johansson, Magnus; Permert, Johan; Karlsson, Anna

doi:10.1074/jbc.273.24.14707

Cited by 56 publications

(32 citation statements)

References 17 publications

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“…A low but significant activity with ganciclovir was also observed with the recombinant enzyme (24), similar to what was reported earlier with native dGK (25). This low activity may be involved in the toxicity observed with ganciclovir (26), although it is not clear how a mitochondrial enzyme can be responsible for the toxicity of nucleoside analogs, which supposedly target nuclear DNA synthesis (35).…”

supporting

confidence: 61%

Antiviral Guanosine Analogs as Substrates for Deoxyguanosine Kinase: Implications for Chemotherapy

Sjöberg

Wang

Eriksson

2001

Antimicrob Agents Chemother

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A highly active form of human recombinant deoxyguanosine kinase (dGK) phosphorylated purine nucleoside analogs active against cytomegalovirus, hepatitis B virus, and human immunodeficiency virus, such as penciclovir, 2,3-dideoxyguanosine and 3-fluoro-2,3-dideoxyguanosine. The antiherpesvirus drug ganciclovir, which is also used in gene therapy, was a substrate for dGK, but with low efficiency. ATP and UTP were both good phosphate donors, with apparent K m values of 6 and 4 M and V max values of 34 and 90 nmol of dGMP/ mg of dGK/min, respectively. With a mixture of 5 mM ATP and 0.05 mM UTP, which represent physiologically relevant concentrations, the activities of dGK with ganciclovir and penciclovir was 1% and approximately 10%, respectively, of that with dGuo. The levels of dGK in different tissues were determined with a selective enzyme assay and the total activities per gram of tissues were similar in liver, brain, heart, and thymus extracts. The fact that the cellular dGK enzyme can phosphorylate antiviral guanosine analogs may help to explain the efficacies and side effects of several forms of chemotherapy.

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supporting

confidence: 61%

Antiviral Guanosine Analogs as Substrates for Deoxyguanosine Kinase: Implications for Chemotherapy

Sjöberg

Wang

Eriksson

2001

Antimicrob Agents Chemother

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“…The mutant NDP kinase could then play the role of a suicide enzyme in case of uncontrolled proliferation (31,32). The overexpression of the mitochondrial deoxyguanosine kinase in human pancreatic adenocarcinoma cell lines was reported to enhance sensitivity of the cells to CdA, araG, and dFdG (33). Promising results have already been obtained by transfection with herpes simplex thymidine kinase that increases the cells sensitivity to ganciclovir (34).…”

Section: Fig 4 Structure Of Ei-n119s In Complex With Rb-azt-tpmentioning

confidence: 58%

Improving Nucleoside Diphosphate Kinase for Antiviral Nucleotide Analogs Activation

Gallois-Montbrun

Schneider

Chen

et al. 2002

Journal of Biological Chemistry

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Nucleotide analogs such as dideoxynucleosides, AZT, 1 and d4T are widely used in clinics for their antiviral effects, in particular in the treatment of AIDS. Because the sugar moiety of these nucleoside reverse transcriptase inhibitors (NRTI) lacks a 3Ј-OH group, their incorporation by viral DNA polymerase or reverse transcriptase leads to DNA chain termination. To be substrates of DNA synthesis, an analog must first be converted to the 5Ј-triphosphate form, which is done intracellularly by kinases of the nucleoside salvage pathway. Whereas the first two phosphorylation steps are catalyzed by enzymes specific for the nucleobase, the ␥-phosphate is added by nucleoside diphosphate (NDP) kinase, which exhibits little specificity toward the nucleobase and the sugar moiety (1). The ␥-phosphate transfer from N 1 TP to N 2 DP catalyzed by NDP kinase involves a phosphohistidine intermediate,All eukaryotic NDP kinases are homohexamers with a 17-kDa subunit (2). In humans, where eight isoforms have been reported, the major isoforms NDPK-A and NDPK-B, respectively encoded by the genes nm23-H1 and nm23-H2, display 88% sequence identity and have very similar kinetic parameters (3). They closely resemble the NDP kinase from the lower eukaryote Dictyostelium discoideum (Dd-NDPK), which for most purposes is as a reliable model of other eukaryotic NDP kinases (4), easier to purify and crystallize than human NDP kinases. NDP kinases have a very high turnover rate on natural nucleotides, but their catalytic efficiency drops by a factor of 10 4 on the analogs AZT diphosphate or ddNDP (5). This is attributed to the substrate-assisted catalysis mechanism of NDP kinase, where the 3Ј-OH plays a major role. Using fluorescence stopped-flow experiments to study the two half-reactions (Reactions 1 and 2), we have previously shown that affinity is reduced 10-fold and phosphotransfer 500-to 1000-fold slower in the absence of the 3Ј-OH (6). The poor activation of NRTI by NDP kinase, resulting in low amounts of the triphosphate form of NRTI within infected cells, is of clinical importance. It is a major cause of incomplete suppression of viral DNA synthesis, allowing the selection of resistance mutations (7).To overcome this limitation, we designed new NRTIs with increased reactivity toward NDP kinase: the ␣-borano deriva-

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“…A possible explanation is that up-or down-regulation of nucleoside metabolizing enzymes in our HEK293 cells limits the potential for transport by MRP4 or MRP5. On the one hand, the toxicity of gemcitabine, cytarabine, and cladribine in cell lines increases proportionally with deoxycytidine kinase activity (Galmarini et al, 2001), and transfection with mitochondrial deoxyguanosine kinase has also been shown to increase sensitivity to the cytostatic activity of these agents (Zhu et al, 1998). On the other hand, transfection with human cytosolic nucleotidase imparts HEK293 cells with resistance against cladribine and gemcitabine (Hunsucker et al, 2001), and we have previously found that thiopurine nucleotides were rapidly dephosphorylated in HEK293 cells, limiting the toxicity of thiopurine drugs (Wielinga et al, 2002).…”

Section: Discussionmentioning

confidence: 99%

Characterization of the Transport of Nucleoside Analog Drugs by the Human Multidrug Resistance Proteins MRP4 and MRP5

Reid

Wielinga²,

Zelcer³

et al. 2003

Mol Pharmacol

339

259

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The human multidrug resistance proteins MRP4 and MRP5 are organic anion transporters that have the unusual ability to transport cyclic nucleotides and some nucleoside monophosphate analogs. Base and nucleoside analogs used in the chemotherapy of cancer and viral infections are potential substrates. To assess the possible contribution of MRP4 and MRP5 to resistance against these drugs, we have investigated the transport mediated by MRP4 and MRP5. In cytotoxicity assays, MRP4 conferred resistance to the antiviral agent 9-(2-phosphonomethoxyethyl)adenine (PMEA) and high-performance liquid chromatography analysis showed that, like MRP5, MRP4 transported PMEA in an unmodified form. MRP4 also mediated substantial resistance against other acyclic nucleoside phosphonates, whereas MRP5 did not. Apart from lowlevel MRP4-mediated cladribine resistance, the cytotoxicity of clinically used anticancer nucleosides was not influenced by overexpression of MRP4 or MRP5. In contrast, MRP5 mediated efflux of the pyrimidine-based antiviral 2Ј,3Ј-dideoxynucleoside 2Ј,3Ј-didehydro-2Ј,3Ј-dideoxythymidine 5Ј-monophosphate (d4TMP) and its phosphoramidate derivative alaninyl-d4TMP from cells loaded with the 2Ј,3Ј-didehydro-2Ј,3Ј-dideoxythymidine prodrugs cyclosaligenyl-d4TMP and aryloxyphosphoramidate d4TMP (So324), respectively. Moreover, only inside-out membrane vesicles derived from MRP5-overexpressing cells accumulated alaninyl-d4TMP. Cellular efflux and vesicular uptake studies were carried out to further compare transport mediated by MRP4 and MRP5 and showed that dipyridamole, dilazep, nitrobenzyl mercaptopurine riboside, sildenafil, trequinsin and MK571 inhibited MRP4 more than MRP5, whereas cyclic nucleotides and monophosphorylated nucleoside analogs were equally poor inhibitors of both pumps. These results strongly suggest that the affinity of MRP4 and MRP5 for nucleotide-based substrates is low.

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Enhanced Cytotoxicity of Nucleoside Analogs by Overexpression of Mitochondrial Deoxyguanosine Kinase in Cancer Cell Lines

Cited by 56 publications

References 17 publications

Antiviral Guanosine Analogs as Substrates for Deoxyguanosine Kinase: Implications for Chemotherapy

Antiviral Guanosine Analogs as Substrates for Deoxyguanosine Kinase: Implications for Chemotherapy

Improving Nucleoside Diphosphate Kinase for Antiviral Nucleotide Analogs Activation

Characterization of the Transport of Nucleoside Analog Drugs by the Human Multidrug Resistance Proteins MRP4 and MRP5

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