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Nucleic Acids Research, 2000, Vol. 28, No. 5 1170-1175
© 2000 Oxford University Press

Stereochemical control of DNA biosynthesis

Vasily V. Sosunov1, Fanny Santamaria2, Lyubov S. Victorova1,3, Gilles Gosselin2, Bernard Rayner2,* and Alexander A. Krayevsky1

1Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 32 Vavilov Str., Moscow 117984, Russia, 2Laboratoire de Chimie Organique Biomoléculaire de Synthèse, UMR 5625 CNRS-UM II, Université Montpellier II, Place Eugène Bataillon, 34095 Montpellier Cedex 5, France and 3Centre for Medical Studies of University of Oslo, 32 Vavilov Str., Moscow 117984, Russia

Stereochemical control of DNA biosynthesis was studied using several DNA-synthesizing complexes containing, in each case, a single substitution of a 2'-deoxy-D-nucleotide residue by an enantiomeric L-nucleotide residue in a DNA chain (either in the primer or in the template) as well as 2'-deoxy-L-ribonucleoside 5'-triphosphates (L-dNTPs) as substrates. Three template-dependent DNA polymerases were tested, Escherichia coli DNA polymerase I Klenow fragment, Thermus aquaticus DNA polymerase and avian myeloblastosis virus reverse transcriptase, as well as template-independent calf-thymus terminal deoxynucleotidyl transferase. Very stringent control of stereoselectivity was demonstrated for template-dependent DNA polymerases, whereas terminal deoxynucleotidyl transferase was less selective. DNA polymerase I and reverse transcriptase catalyzed formation of dinucleoside 5',5'-tetraphosphates when L-dTTP was used as substrate. Comparison between models of template–primer complexes, modified or not by a single L-nucleotide residue, revealed striking differences in their geometry.

* To whom correspondence should be addressed. Tel: +33 467143394; Fax: +33 467042029; Email: rayner@univ-montp2.fr Alexander A. Krayevsky, deceased August 24, 1999


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