Nucleic Acids Research, 2003, Vol. 31, No. 23 6916-6925
© 2003 Oxford University Press
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Mutagenesis of human DNA polymerase 
: essential roles of Tyr505 and Phe506 for both DNA polymerase and terminal transferase activities
Institute of Veterinary Biochemistry and Molecular Biology, University of Zürich-Irchel, Winterthurerstrasse 190, CH-8057, Zürich, Switzerland, 1 Istituto di Genetica Molecolare, IGM-CNR, Via Abbiategrasso 207, I-27100 Pavia, Italy and 2 Institut de Pharmacologie et de Biologie Structurale, Centre National de la Recherche Scientifique, 205 Route de Narbonne, 310077 Toulouse Cedex, France
*To whom correspondence should be addressed. Tel: +39 382546355; Fax: +39 382422286; Email: maga{at}igm.cnr.it
The authors wish it to be known that, in their opinion, the first two authors should be considered joint First Authors
DNA polymerase (pol) 
is homologous to pol ß and has intrinsic polymerase and terminal transferase activities. However, nothing is known about the amino acid residues involved in these activites. In order to precisely define the nucleotide-binding site of human pol , we have mutagenised two amino acids, Tyr505 and the neighbouring Phe506, which were predicted by structural homology modelling to correspond to the Tyr271 and Phe272 residues of pol ß, which are involved in nucleotide binding. Our analysis demonstrated that pol Phe506Arg/Gly mutants possess very low polymerase and terminal transferase activities as well as greatly reduced abilities for processive DNA synthesis and for carrying on translesion synthesis past an abasic site. The Tyr505Ala mutant, on the other hand, showed an altered nucleotide binding selectivity to perform the terminal transferase activity. Our results suggest the existence of a common nucleotide-binding site for the polymerase and terminal transferase activities of pol , as well as distinct roles of the amino acids Tyr505 and Phe506 in these two catalytic functions.
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