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Research article2003Peer reviewed

Molecular characterization of thymidine kinase from Ureaplasma urealyticum: nucleoside analogues as potent inhibitors of mycoplasma growth

Carnrot, C; Wehelie, R; Eriksson, S; Bolske, G; Wang, LY

Abstract

Ureaplasma urealyticum (U. urealyticum), belonging to the class Mollicutes, is a human pathogen colonizing the urogenital tract and causes among other things respiratory diseases in premature infants. We have studied the salvage of pyrimidine deoxynucleosides in U. urealyticum and cloned a key salvage enzyme, thymidine kinase (TK) from U. urealyticum. Recombinant Uu-TK was expressed in E. coli, purified and characterized with regards to substrate specificity and feedback inhibition. Uu-TK efficiently phosphorylated thymidine (dThd) and deoxyuridine (dUrd) as well as a number of pyrimidine nucleoside analogues. All natural ribonucleoside/deoxyribonucleoside triphosphates, except dTTP, served as phosphate donors, while dTTP was a feedback inhibitor. The level of Uu-TK activity in U. urealyticum extracts increased upon addition of dUrd to the growth medium. Fluoropyrimidine nucleosides inhibited U. urealyticum and M. pneumoniae growth and this inhibitory effect could be reversed by addition of dThd, dUrd or deoxytetrahydrouridine to the growth medium. Thus, the mechanism of inhibition was most likely the depletion of dTTP, either via a blocked thymidine kinase reaction and/or thymidylate synthesis step and these metabolic reactions should be suitable targets for antimycoplasma chemotherapy.

Published in

Molecular Microbiology
2003, Volume: 50, number: 3, pages: 771-780
Publisher: BLACKWELL PUBLISHING LTD

      SLU Authors

    • Eriksson, Staffan

      • Department of Molecular Biosciences, Swedish University of Agricultural Sciences

      • Carnrot, Cecilia

        • Department of Molecular Biosciences, Swedish University of Agricultural Sciences

        • Wehelie, Rahma

          • Department of Molecular Biosciences, Swedish University of Agricultural Sciences

          • Wang, Liya

            • Department of Molecular Biosciences, Swedish University of Agricultural Sciences

          UKÄ Subject classification

          Cell and Molecular Biology
          Microbiology in the medical area

          Publication identifier

          DOI: https://doi.org/10.1046/j.1365-2958.2003.03717.x

          Permanent link to this page (URI)

          https://res.slu.se/id/publ/1817