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Nucleic Acids Research, 2003, Vol. 31, No. 6 1665-1672
© 2003 Oxford University Press

Mosquito has a single multisubstrate deoxyribonucleoside kinase characterized by unique substrate specificity

Wolfgang Knecht1, Gitte Ebert Petersen1,2, Michael Paolo Bastner Sandrini1, Leif Søndergaard3, Birgitte Munch-Petersen4 and Jure Piskur1

1 Eukaryote Molecular Biology Group, BioCentrum-DTU, Building 301, Technical University of Denmark, DK 2800 Lyngby, Denmark, 2 Jubile Kinase ApS, Bavnebjergspark 36, DK 3520 Farum, Denmark, 3 Department of Genetics, Copenhagen University, DK 1353 Copenhagen, Denmark and 4 Department of Life Sciences and Chemistry, Roskilde University, PO Box 260, DK 4000 Roskilde, Denmark

*To whom correspondence should be addressed. Tel: +45 45252518; Fax: +45 45932809; Email: jp{at}biocentrum.dtu.dk
Present address:
Wolfgang Knecht, Astra Zeneca R&D Moelndal, S 43183 Moelndal, Sweden

In mammals four deoxyribonucleoside kinases, with a relatively restricted specificity, catalyze the phosphorylation of the four natural deoxyribonucleosides. When cultured mosquito cells, originating from the malaria vector Anopheles gambiae, were examined for deoxyribonucleoside kinase activities, only a single enzyme was isolated. Subsequently, the corresponding gene was cloned and over-expressed. While the mosquito kinase (Ag-dNK) phosphorylated all four natural deoxyribonucleosides, it displayed an unexpectedly higher relative efficiency for the phosphorylation of purine versus pyrimidine deoxyribonucleosides than the fruit fly multisubstrate deoxyribonucleoside kinase (EC 2.7.1.145). In addition, Ag-dNK could also phosphorylate some medically interesting nucleoside analogs, like stavudine (D4T), 2-chloro-deoxyadenosine (CdA) and 5-bromo-vinyl-deoxyuridine (BVDU). Although the biological significance of multisubstrate deoxyribonucleoside kinases and their diversity among insects remains unclear, the observed variation provides a whole range of applications, as species specific and highly selective targets for insecticides, they have a potential to be used in the enzymatic production of various (di-)(deoxy-)ribonucleoside monophosphates, and as suicide genes in gene therapy.


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