title: Mycoplasma pneumoniae thymidine phosphorylase.
abstract: Mycoplasma pneumoniae (Mpn) is a human pathogen causing acute respiratory diseases and accounts for approximately 30% cases of community-acquired pneumonia. Co-infection with Mycoplasmas compromises the efficacy of anticancer and antiviral nucleoside analog-based drugs due to the presence of Mycoplasma thymidine phosphorylase (TP). In this study, a TP-deficient strain of Mpn was generated in order to study the effect of Mpn TP in the metabolism of nucleoside analogs. Deficiency in TP activity led to increased uptake and incorporation of radiolabeled deoxyuridine and uracil but thymidine uptake was not affected. The activities of enzymes in the salvage of thymidine and deoxyuridine, e.g., thymidine kinase and uracil phosphoribosyltransferase were upregulated in the TP-deficient mutant, which may explain the increased uptake of deoxyuridine and uracil. Thirty FDA-approved anticancer and antiviral nucleoside and nucleobase analogs were used to screen their inhibitory activity toward the TP mutant and the wild type strain. Seven analogs were found to inhibit strongly the growth of both wild type and TP mutant. Differences in the inhibitory effect of several purine analogs between the two strains were observed. Further study is needed in order to understand the mechanism of inhibition caused by these analogs. Our results indicated that TP is not an essential gene for Mpn survival and TP deficiency affects other enzymes in Mpn nucleotide metabolism, and suggested that Mycoplasma nucleotide biosynthesis pathway enzymes are potential targets for future development of antibiotics.
authors.affiliation: a Department of Anatomy, Physiology and Biochemistry , Swedish University of Agricultural Sciences, The Biomedical Center , Uppsala , Sweden.
authors.forename: Liya
authors.initials: L
authors.lastname: Wang
authors.forename: Sebastian R
authors.initials: SR
authors.lastname: Schmidl
authors.forename: Jörg
authors.initials: J
authors.lastname: Stülke
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology; D000998:Antiviral Agents / Q000494:pharmacology; D003857:Deoxyuridine / Q000378:metabolism; D009154:Mutation; D009177:Mycoplasma pneumoniae / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D013936:Thymidine / Q000378:metabolism; D013939:Thymidine Phosphorylase / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*
chemical_list: D000970:Antineoplastic Agents; D000998:Antiviral Agents; D013939:Thymidine Phosphorylase; D013936:Thymidine; D003857:Deoxyuridine
keywords: Mycoplasma pneumonia; growth inhibition; nucleoside and nucleobase analogs; thymidine kinase; thymidine phosphorylase