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. 2014 Sep 27;42(19):12102–12111. doi: 10.1093/nar/gku879

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© The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Processivity of human PrimPol and PrimPol^Y89D. Primer extension assays were performed in the presence of an excess of herring sperm DNA trap to ensure the polymerase only binds once to the template DNA. Control reactions were also carried out to test the effectiveness of the trap DNA (data not shown). The first lanes represent a no-enzyme control and lanes 2–6 represent time points of 15, 30, 60, 120 and 360 s. (A) Human PrimPol is a poorly processive enzyme, inserting up to four nucleotides opposite a templating DNA strand in a single binding event. (B) PrimPol^Y89D has marked reduction in processivity, and is only able to insert a single nucleotide opposite a templating DNA strand per single binding event. (C) PrimPol^Y89S can also insert only one single nucleotide opposite a templating DNA strand. (D) PrimPol^Y89F restores processivity and has the ability to insert up to four nucleotides, similarly to wild-type PrimPol.