Human deoxycytidine kinase is located in the cell nucleus

Johansson, Magnus; Brismar, Sophia; Karlsson, Anna

doi:10.1073/pnas.94.22.11941

Cited by 72 publications

(63 citation statements)

References 36 publications

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“…We found no difference in either enzyme activity, cellular sensitivity to nucleoside analogs, or bystander cell killing when the enzyme was expressed in the cytosol or in the nucleus. These results are consistent with a model of rapid equilibration of the dNTP pools between the nucleus and the cytosol mediated by the nuclear pore complexes (Johansson et al, 1997;Zhu et al, 2000).…”

supporting

confidence: 80%

Mitochondrial Expression of theDrosophila melanogasterMultisubstrate Deoxyribonucleoside Kinase

Solaroli

Zheng²,

Johansson³

et al. 2007

Mol Pharmacol

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The multisubstrate deoxyribonucleoside kinase from Drosophila melanogaster deoxyribonucleoside kinase (Dm-dNK) is studied as a candidate suicide gene for applications in combined gene/chemotherapy of cancer. We have created an engineered Dm-dNK nucleoside kinase that is targeted to the mitochondrial matrix. The enzyme was expressed in a thymidine kinase 1-deficient osteosarcoma cell line, and the sensitivity of the cells to cytotoxic nucleoside analogs was determined when the enzyme was targeted to either the nucleus or the mitochondrial matrix. Although the total deoxythymidine (dThd) phosphorylation activity was similar in cells expressing Dm-dNK in the nucleus or in the mitochondria, the cells expressing the enzyme in the mitochondria showed higher sensitivity to the antiproliferative activity of several pyrimidine nucleoside analogs, such as (E)-5-(2-bromovinyl)-2Ј-deoxyuridine, 5-bromo-2Ј-deoxyuridine, and 5-fluoro-2Ј-deoxyuridine. Labeling studies using [ 3 H]dThd showed that the cells expressing the mitochondrial enzyme had an increased incorporation of [ 3 H]dThd into DNA, shown to be due to a higher [3 H]dTTP specific activity of the total dTTP pool in the cells in which Dm-dNK was targeted to the mitochondria. The difference in the specific activity of the dTTP pool is a result of different contributions of the de novo and the salvage pathways for the dTTP synthesis in transduced cells. In summary, these findings suggest that mitochondrial targeting of Dm-dNK facilitates nucleoside and nucleoside analog phosphorylation and could be used as a strategy to enhance the efficacy of nucleoside analog phosphorylation and concomitantly their cytostatic potential.

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supporting

confidence: 80%

Mitochondrial Expression of theDrosophila melanogasterMultisubstrate Deoxyribonucleoside Kinase

Solaroli

Zheng²,

Johansson³

et al. 2007

Mol Pharmacol

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“…2D). Since the nucleoside analogs may freely traverse the nuclear envelope, phosphorylating in either of the two subcellular compartments (25). The recombinant dNKmut was more favorable for medical therapy.…”

Section: Discussionmentioning

confidence: 99%

Adenovirus-mediated Drosophila melanogaster deoxyribonucleoside kinase mutants combined with gemcitabine harbor a safe cancer treatment profile

Zhu

Ma²,

Zhao³

et al. 2011

Int J Oncol

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Abstract. The purpose of this analysis was to investigate the enzyme activity and specificity of adenovirus-mediated Drosophila melanogaster deoxyribonucleoside kinase (DmdNK) mutants in combination with gemcitabine. Compared to herpes simplex type 1 thymidine kinases (HSV-TK) and other known dNKs, this Dm-dNK enzyme has a broader substrate specificity and a higher catalytic rate. We created the Dm-dNK mutants (dNKmut) by site-directed mutagenesis at the sites of 244E, 245S, 251S, and 252R, with the last 10 amino acids in the amino acid sequence randomly alternated. We subsequently evaluated the enzyme activity and substrate specificity. The engineered enzymes showed a relative increase in phosphorylation in the nucleoside analogs of gemcitabine (dFdC, 2',2'-difluoro-deoxycytidine) compared to the wild-type enzyme. The dNKmut enzymes were expressed in breast (Bcap37) and gastric (SGC-7901) cancer cell lines. In studying the sensitivity of the cell lines to dFdC, conditionally replicative adenovirus (CRAd) ZD55-dNKmut showed higher expression and enzymatic activity than the replication-defective adenovirus Ad-dNKmut in cancer cells, but with less cytotoxicity to cancer cells than that of Ad-dNKmut. Our data suggest that the triple phosphorylated dFdC catalyzed by dNKmut inhibited the replication of adenovirus with a simultaneous positive therapeutic effect on cancer cells. Therefore, concomitant use of the ZD55-dNKmut and dFdC could be a novel targeted strategy in suicide gene therapy with safe control of excessive virus replication.

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“…Cells lacking hENT1 are highly resistant to gemcitabine (36,37), and pancreas cancer patients with hENT1-positive tumor tissue have significantly longer survival after gemcitabine chemotherapy than patients without detectable hENT1 (38). dCK plays a key role in the activation of gemcitabine, and its activity correlates with the drug sensitivity (39)(40)(41). Deficiency in the dCK activity has been considered to be one of the main mechanisms for the development of resistance to gemcitabine (42).…”

Section: Introductionmentioning

confidence: 99%

Ribonucleotide reductase subunit M1 assessed by quantitative double-fluorescence immunohistochemistry predicts the efficacy of gemcitabine in biliary tract carcinoma

Nakamura

Kohya²,

Kai³

et al. 2010

Int J Oncol

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Abstract. Gemcitabine is a commonly used chemotherapeutic agent for advanced biliary tract carcinoma (BTC), although its efficacy is insufficient. Therefore, it is essential to establish new diagnostic methods, which can predict responders before the treatment. The aim of this study is to identify the most reliable chemoresistance marker to gemcitabine in BTC among the 4 molecules (hENT1, dCK, RRM1 and RRM2) involved in gemcitabine metabolism. The expression of 4 molecules were investigated in 5 BTC cell lines, and correlated with gemcitabine sensitivity. RRM1 protein was also assessed by quantitative double-fluorescence immunohistochemistry (qDFIHC) in 10 patients with unresectable or recurrent BTC who received gemcitabine-based chemotherapy. RRM1 and RRM2 protein strongly correlated with the IC 50 value for gemcitabine in BTC cell lines (R=0.935, 0.771, respectively). In addition, patients with low RRM1 were significantly more sensitive to gemcitabine (p=0.033), and their survival was significantly better than patients with high RRM1 (p=0.001).In conclusion, RRM1 particularly in protein level is a reliable marker for gemcitabine resistance in BTC. Furthermore, qDFIHC is a useful method for the assessment of RRM1 protein, in order to design a tailor-made chemotherapeutic regimen for BTC patients.

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Human deoxycytidine kinase is located in the cell nucleus

Cited by 72 publications

References 36 publications

Mitochondrial Expression of theDrosophila melanogasterMultisubstrate Deoxyribonucleoside Kinase

Mitochondrial Expression of theDrosophila melanogasterMultisubstrate Deoxyribonucleoside Kinase

Adenovirus-mediated Drosophila melanogaster deoxyribonucleoside kinase mutants combined with gemcitabine harbor a safe cancer treatment profile

Ribonucleotide reductase subunit M1 assessed by quantitative double-fluorescence immunohistochemistry predicts the efficacy of gemcitabine in biliary tract carcinoma

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