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A more recent version of this article appeared on August 10, 2001
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M105272200v1
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Papers In Press, published online ahead of print June 13, 2001
J. Biol. Chem, 10.1074/jbc.M105272200
Submitted on June 7, 2001
Revised on June 13, 2001
Accepted on June 12, 2001

Terminal deoxynucleotidyl transferase indiscriminately incorporates ribonucleotides and deoxyribonucleotides

Jean-Baptiste Boulé, François Rougeon, and Catherine Papanicolaou

Immunologie, Institut Pasteur, Paris 75015

Corresponding Author: papanico{at}pasteur.fr

Terminal deoxynucleotidyl Transferase (TdT) catalyzes the condensation of deoxyribonucleotides on 3'-hydroxyl ends of DNA strands in a template-independent manner and adds N-regions to gene segment junctions during V(D)J recombination. Although TdT is able to incorporate a few ribonucleotides in vitro, TdT discrimination between ribo and deoxyribonucleotides was never studied. We found that TdT shows only minor preference for incorporation of deoxyribonucleotides over ribonucleotides on DNA strands. However, incorporation of ribonucleotides alone or in the presence of deoxyribonucleotides generally leads to premature chain termination, reflecting an impeded accommodation of ribo or mixed ribo/deoxyribo nucleic acid substrates by TdT. An essential catalytic aspartate in TdT was identified, a first step toward understanding the apparent lack of sugar discrimination by TdT.


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