Toggle Main Menu Toggle Search

Open Access padlockePrints

Recognition of the Pro-mutagenic Base Uracil by Family B DNA Polymerases from Archaea

Lookup NU author(s): Gillian Shuttleworth, Professor Bernard Connolly


Full text for this publication is not currently held within this repository. Alternative links are provided below where available.


Archaeal family B DNA polymerases contain a specialised pocket that binds tightly to template-strand uracil, causing the stalling of DNA replication. The mechanism of this unique "template-strand proof-reading" has been studied using equilibrium binding measurements, DNA footprinting, van't Hoff analysis and calorimetry. Binding assays have shown that the polymerase preferentially binds to uracil in single as opposed to double-stranded DNA. Tightest binding is observed using primer-templates that contain uracil four bases in front of the primer-template junction, corresponding to the observed stalling position. Ethylation interference analysis of primer-templates shows that the two phosphates, immediately flanking the uracil (NpUpN), are important for binding; contacts are also made to phosphates in the primer-strand. Microcalorimetry and van't Hoff analysis have given a fuller understanding of the thermodynamic parameters involved in uracil recognition. All the results are consistent with a "read-ahead" mechanism, in which the replicating polymerase scans the template, ahead of the replication fork, for the presence of uracil and halts polymerisation on detecting this base. Post-stalling events, serving to eliminate uracil, await full elucidation. © 2004 Elsevier Ltd. All rights reserved.

Publication metadata

Author(s): Shuttleworth G, Fogg MJ, Kurpiewski MR, Jen-Jacobson L, Connolly BA

Publication type: Article

Publication status: Published

Journal: Journal of Molecular Biology

Year: 2004

Volume: 337

Issue: 3

Pages: 621-634

ISSN (print): 0022-2836

ISSN (electronic): 1089-8638

Publisher: Academic Press


DOI: 10.1016/j.jmb.2004.01.021

PubMed id: 15019782


Altmetrics provided by Altmetric