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Nucleic Acids Research 2005 33(13):4117-4127; doi:10.1093/nar/gki723
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Published online 25 July 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oupjournals.org

Article

Incorporation of non-nucleoside triphosphate analogues opposite to an abasic site by human DNA polymerases ß and {lambda}

Emmanuele Crespan, Samantha Zanoli, Anastasiya Khandazhinskaya1, Igor Shevelev2, Maxim Jasko1, Ludmila Alexandrova1, Marina Kukhanova1, Giuseppina Blanca3, Giuseppe Villani3, Ulrich Hübscher2, Silvio Spadari and Giovanni Maga*

Istituto di Genetica Molecolare IGM-CNR via Abbiategrasso 207, I-27100 Pavia, Italy 1Engelhardt Institute of Molecular Biology, RAS 32 Vavilov Street, 119991 Moscow, Russian Federation, Russia 2Institute of Veterinary Biochemistry and Molecular Biology University of Zürich–Irchel Winterthurerstrasse 190, CH-8057 Zürich, Switzerland 3Institut de Pharmacologie et de Biologie Structurale, Centre National de la Recherche Scientifique 205 route de Narbonne, 31077 Toulouse Cedex, France

*To whom correspondence should be addressed. Tel: +39 0382546354; Fax: +39 0382422286; Email: maga{at}igm.cnr.it

Received June 9, 2005. Revised July 5, 2005. Accepted July 5, 2005.

A novel class of non-nucleoside triphosphate analogues, bearing hydrophobic groups sterically similar to nucleosides linked to the {alpha}-phosphate but lacking the chemical functional groups of nucleic acids, were tested against six different DNA polymerases (polymerases). Human polymerases {alpha}, ß and {lambda}, and Saccharomyces cerevisiae polymerase IV, were inhibited with different potencies by these analogues. On the contrary, Escherichia coli polymerase I and HIV-1 reverse transcriptase were not. Polymerase ß incorporated these derivatives in a strictly Mn++-dependent manner. On the other hand, polymerase {lambda} could incorporate some alkyltriphosphate derivatives with both Mg++ and Mn++, but only opposite to an abasic site on the template strand. The active site mutant polymerase {lambda} Y505A showed an increased ability to incorporate the analogues. These results show for the first time that neither the base nor the sugar moieties of nucleotides are required for incorporation by family X DNA polymerases.

The authors wish to be known that, in their opinion, the first two authors should be regarded as joint First Authors


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