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. 2011 Nov 15;40(5):2089–2106. doi: 10.1093/nar/gkr983

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© The Author(s) 2011. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 8. — Mph1 DNA helicase activity on replication fork substrates. (A) Helicase activity of Mph1 on fixed or partially movable double-stranded three-way junction substrates. Mph1 (120 fmol) was incubated using reaction conditions in Figure 6. (B) Replication fork regression activity of Mph1 on completely movable double-stranded three-way junction substrate. Assays were performed with 60 fmol of Mph1 using standard reaction conditions (20 μl) for various incubation periods (0, 1, 2, 5, 15 and 30 min). The amount of total product (slower and faster migrating band) formed by regression is indicated below each lane. In this case, the level of product formed in the absence of Mph1 was not subtracted. (C) Illustration how two dsDNA products are formed by regression of replication forks via chicken-foot structure. (D) Replication fork regression activity of Mph1 with various ribo- and deoxyribonucleotides as cofactors. Mph1 (50 fmol) was incubated using standard reaction condition for 15 min and the amount of product formed is shown as a bar graph. The level of product formed in the absence of Mph1 was subtracted from the values presented.