Abstract
In the Pasteurella multocida genome only one putative deoxyribonucleoside kinase encoding gene, for thymidine kinase 1 (PmTK1), was identified. The PmTK1 gene was sub-cloned into Escherichia coli KY895 and it sensitized the host towards 2',2'-difluoro-deoxycytidine (gemcitabine, dFdC), 3'-azido-thymidine (AZT) and 5-fluoro-deoxyuridine (5F-dU). PmTK1 was over-expressed and purified with two different tags. Apparently, deoxyuridine (dU), and not thymidine (dT), is the preferred substrate. We suggest that PmTK1s could be employed as a species-specific activator of uracil-based nucleoside antibiotics.
MeSH terms
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Antimetabolites, Antineoplastic / metabolism
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Antimetabolites, Antineoplastic / pharmacology
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Deoxycytidine / analogs & derivatives
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Deoxycytidine / metabolism
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Deoxycytidine / pharmacology
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Deoxyuridine / pharmacology
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Escherichia coli / drug effects
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Gemcitabine
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Microbial Sensitivity Tests
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Pasteurella multocida / enzymology*
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Pyrimidine Nucleosides / metabolism*
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Pyrimidine Nucleosides / pharmacology
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Thymidine Kinase / genetics
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Thymidine Kinase / metabolism*
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Zidovudine / pharmacology
Substances
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Antimetabolites, Antineoplastic
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Pyrimidine Nucleosides
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Deoxycytidine
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Zidovudine
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Thymidine Kinase
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thymidine kinase 1
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Deoxyuridine
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Gemcitabine