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. 2014 Mar 12;42(9):5657–5670. doi: 10.1093/nar/gku198

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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-nc/3.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 7. — Enhancement of FANCI ubiquitination by DNA. (A) Various forms of ϕX plasmid DNA (11 or 45 nM; HIII-HaeIII digested) were added to ubiquitination reactions that contain FANCI (I) or the FANCI K523R mutant. The position of monoubiquitinated FANCI (I-Ub) is marked. The percent of ubiquitinated FANCI product (% Ub) is indicated. (B) FANCI ubiquitination reactions were carried out with linear dsDNA (ds), replication fork (RF) DNA or ss oligonucleotides (O1, O2 and O3) that differ in sequence and length. Reaction products were analyzed by SDS-PAGE and fluorescent staining (18-h incubation) or western blot (3-h incubation). Reaction product formed after 18 h of incubation was quantified. (C) FANCI ubiquitination reactions were performed as in panel (A) but comparing two splayed-arm DNA substrates that differ in length.