Terminal deoxynucleotidyl transferase indiscriminately incorporates ribonucleotides and deoxyribonucleotides.

Abstract:

Terminal deoxynucleotidyl transferase (TdT) catalyzes the condensation ...
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 has never been studied. We found that TdT shows only a 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/deoxyribonucleic acid substrates by TdT. An essential catalytic aspartate in TdT was identified, which is a first step toward understanding the apparent lack of sugar discrimination by TdT.

Polymerases:

Topics:

Kinetic Parameters, Source / Purification

Status:

new topics/pols set partial results complete validated

Results:

Polymerase Reference Property Result Context
Terminal Transferase Terminal deoxynucleotidyl transferase indiscriminately incorporates ribonucleotides and deoxyribonucleotides. Full length or truncated Full length
Terminal Transferase Terminal deoxynucleotidyl transferase indiscriminately incorporates ribonucleotides and deoxyribonucleotides. KM 300uM Reaction: Nucleotide incorporation; Substrate: dATP
Terminal Transferase Terminal deoxynucleotidyl transferase indiscriminately incorporates ribonucleotides and deoxyribonucleotides. KM 368uM Reaction: Nucleotide incorporation; Substrate: rATP
Terminal Transferase Terminal deoxynucleotidyl transferase indiscriminately incorporates ribonucleotides and deoxyribonucleotides. KM 15.9uM Reaction: Nucleotide incorporation; Substrate: dATP
Terminal Transferase Terminal deoxynucleotidyl transferase indiscriminately incorporates ribonucleotides and deoxyribonucleotides. KM 5.9uM Reaction: Nucleotide incorporation; Substrate: dATP
Terminal Transferase Terminal deoxynucleotidyl transferase indiscriminately incorporates ribonucleotides and deoxyribonucleotides. kcat 76 /minute Reaction: Nucleotide incorporation; Substrate: rATP
Terminal Transferase Terminal deoxynucleotidyl transferase indiscriminately incorporates ribonucleotides and deoxyribonucleotides. kcat 180 /minute Reaction: Nucleotide incorporation; Substrate: dATP
Terminal Transferase Terminal deoxynucleotidyl transferase indiscriminately incorporates ribonucleotides and deoxyribonucleotides. kcat 147 /minute Reaction: Nucleotide incorporation; Substrate: dATP
Terminal Transferase Terminal deoxynucleotidyl transferase indiscriminately incorporates ribonucleotides and deoxyribonucleotides. kcat 11.3 /minute Reaction: Nucleotide incorporation; Substrate: dATP

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