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Nucleic Acids Research, 2001, Vol. 29, No. 11 2217-2222
© 2001 Oxford University Press

DNA synthesis fidelity by the reverse transcriptase of the yeast retrotransposon Ty1

Mansour Boutabout, Marcelle Wilhelm and François-Xavier Wilhelm*

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

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 At:A mispair in which dATP is misincorporated opposite a template A to 6.27 x 10–5 for the Ct:A mispair. The Gt:G and Tt: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 ~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.

* To whom correspondence should be addressed. Tel: +33 3 88 41 70 06; Fax: +33 3 88 60 22 18; Email: wilhelm{at}ibmc.u-strasbg.fr


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