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Nucleic Acids Research Advance Access published online on December 5, 2006
Nucleic Acids Research, doi:10.1093/nar/gkl1016
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© 2006 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Nucleic Acid Enzymes

Expanding the repertoire of DNA polymerase substrates: template-instructed incorporation of non-nucleoside triphosphate analogues by DNA polymerases ß and {lambda}

Emmanuele Crespan, Ludmila Alexandrova1, Anastasiya Khandazhinskaya1, Maxim Jasko1, Marina Kukhanova1, Giuseppe Villani2, Ulrich Hübscher3, Silvio Spadari and Giovanni Maga*

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

*To whom correspondence should be addressed. Tel: +39 03825 46354; Fax: +39 03824 22286; Email: maga{at}igm.cnr.it

Received September 5, 2006. Revised November 2, 2006. Accepted November 2, 2006.

We have recently shown that neither the base nor the sugar moieties of a nucleotide is an essential feature for its incorporation by DNA polymerases (pols) {lambda} and ß. Here we present the identification of novel non-nucleoside triphosphate (NNTP) derivatives belonging to three classes: (i) non-substrate-specific inhibitors of DNA pol {lambda}; (ii) substrate inhibitors which could preferentially be incorporated by either DNA pol {lambda} wild type or its Y505A mutant and (iii) the substrate inhibitor N-(Biphenylcarbonyl)-4-oxobutyl triphosphate which could be incorporated exclusively by DNA pol ß in a Mg2+-dependent manner, and preferentially pairs with A on the template. This compound represents the first example of a substrate lacking both nucleobase and ribose residue, showing distinct base-pairing properties with normal bases. Therefore, this NNTP analog can be considered as the prototype of an entirely novel class of DNA pol substrates.


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E. Crespan, U. Hubscher, and G. Maga
Error-free bypass of 2-hydroxyadenine by human DNA polymerase {lambda} with Proliferating Cell Nuclear Antigen and Replication Protein A in different sequence contexts
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