title: Cellular response to efficient dUTPase RNAi silencing in stable HeLa cell lines perturbs expression levels of genes involved in thymidylate metabolism.
abstract: dUTPase is involved in preserving DNA integrity in cells. We report an efficient dUTPase silencing by RNAi-based system in stable human cell line. Repression of dUTPase induced specific expression level increments for thymidylate kinase and thymidine kinase, and also an increased sensitization to 5-fluoro-2'-deoxyuridine and 5-fluoro-uracil. The catalytic mechanism of dUTPase was investigated for 5-fluoro-dUTP. The 5F-substitution on the uracil ring of the substrate did not change the kinetic mechanism of dUTP hydrolysis by dUTPase. Results indicate that RNAi silencing of dUTPase induces a complex cellular response wherein sensitivity towards fluoropyrimidines and gene expression levels of related enzymes are both modulated.
authors.affiliation: Institute of Enzymology, Hungarian Academy of Sciences, Budapest, Hungary.
authors.forename: Gábor
authors.initials: G
authors.lastname: Merényi
authors.forename: Júlia
authors.initials: J
authors.lastname: Kovári
authors.forename: Judit
authors.initials: J
authors.lastname: Tóth
authors.forename: Enik
authors.initials: E
authors.lastname: Takács
authors.forename: Imre
authors.initials: I
authors.lastname: Zagyva
authors.forename: Anna
authors.initials: A
authors.lastname: Erdei
authors.forename: Beáta G
authors.initials: BG
authors.lastname: Vértessy
mesh_terms: D005467:Floxuridine / Q000378:metabolism; D005472:Fluorouracil / Q000378:metabolism; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D009703:Nucleoside-Phosphate Kinase / Q000235:genetics* / Q000378:metabolism; D011755:Pyrophosphatases / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D013937:Thymidine Kinase / Q000235:genetics* / Q000378:metabolism
chemical_list: D005467:Floxuridine; D013937:Thymidine Kinase; D009703:Nucleoside-Phosphate Kinase; C020093:dTMP kinase; D011755:Pyrophosphatases; C020020:dUTP pyrophosphatase; D005472:Fluorouracil