title: Expression of bovine cytosolic 5'-nucleotidase (cN-II) in yeast: nucleotide pools disturbance and its consequences on growth and homologous recombination.
abstract: Cytosolic 5'-nucleotidase II is a widespread IMP hydrolyzing enzyme, essential for cell vitality, whose role in nucleotide metabolism and cell function is still to be exactly determined. Cytosolic 5'-nucleotidase overexpression and silencing have both been demonstrated to be toxic for mammalian cultured cells. In order to ascertain the effect of enzyme expression on a well-known eukaryote simple model, we expressed cytosolic 5'-nucleotidase II in Saccharomyces cerevisiae, which normally hydrolyzes IMP through the action of a nucleotidase with distinct functional and structural features. Heterologous expression was successful. The yeast cells harbouring cytosolic 5'-nucleotidase II displayed a shorter duplication time and a significant modification of purine and pyrimidine derivatives concentration as compared with the control strain. Furthermore the capacity of homologous recombination in the presence of mutagenic compounds of yeast expressing cytosolic 5'-nucleotidase II was markedly impaired.
authors.affiliation: Dipartimento di Chimica e Farmacia, Università di Sassari, Sassari, Italy. enomis@uniss.it
authors.forename: Simone
authors.initials: S
authors.lastname: Allegrini
authors.forename: Daniela Nicole
authors.initials: DN
authors.lastname: Filoni
authors.forename: Alvaro
authors.initials: A
authors.lastname: Galli
authors.forename: Anita
authors.initials: A
authors.lastname: Collavoli
authors.forename: Rossana
authors.initials: R
authors.lastname: Pesi
authors.forename: Marcella
authors.initials: M
authors.lastname: Camici
authors.forename: Maria Grazia
authors.initials: MG
authors.lastname: Tozzi
mesh_terms: D015720:5'-Nucleotidase / Q000378:metabolism*; D000704:Analysis of Variance; D000818:Animals; D015153:Blotting, Western; D002417:Cattle / Q000378:metabolism*; D002455:Cell Division / Q000502:physiology; D003600:Cytosol / Q000201:enzymology*; D017931:DNA Primers / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D059765:Homologous Recombination / Q000502:physiology*; D009711:Nucleotides / Q000032:analysis*; D012441:Saccharomyces cerevisiae / Q000254:growth & development* / Q000378:metabolism; D013696:Temperature
chemical_list: D017931:DNA Primers; D009711:Nucleotides; D015720:5'-Nucleotidase