title: DNA synthesis fidelity by the reverse transcriptase of the yeast retrotransposon Ty1.
abstract: The fidelity of the yeast retrotransposon Ty1 reverse transcriptase (RT) was determined by an assay based on gel electrophoresis. Steady-state kinetics analyses of deoxyribonucleotide (dNTP) incorporation at a defined primer-template site indicate that Ty1 RT misincorporates dNTP at a frequency of 0.45 x 10(-5) for the A(t):A mispair in which dATP is misincorporated opposite a template A to 6.27 x 10(-5) for the C(t):A mispair. The G(t):G and T(t):T mispairs are formed with very low efficiency. The fidelity parameters of Ty1 RT do not depend on whether RNA or DNA are copied. Relative to lentiviral RTs (HIV-1, HIV-2 or EIAV) Ty1 RT is approximately 10-fold less error prone. Our data also show that the Ty1 RT is able to recapitulate two error-generating mechanisms: extension of mismatches and non-templated addition of nucleotides at the end of a blunt-end primer-template.
authors.affiliation: Unité Propre de Recherche 9002 du Centre National de la Recherche Scientifique, Institut de Biologie Moléculaire et Cellulaire, 15 rue R. Descartes, 67084 Strasbourg, France.
authors.forename: M
authors.initials: M
authors.lastname: Boutabout
authors.forename: M
authors.initials: M
authors.lastname: Wilhelm
authors.forename: F X
authors.initials: FX
authors.lastname: Wilhelm
mesh_terms: D004271:DNA, Fungal / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D009711:Nucleotides / Q000235:genetics / Q000378:metabolism; D012331:RNA, Fungal / Q000235:genetics; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D018626:Retroelements / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D013698:Templates, Genetic
chemical_list: D004271:DNA, Fungal; D009711:Nucleotides; D012331:RNA, Fungal; D018626:Retroelements; D012194:RNA-Directed DNA Polymerase