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. 2019 Jun 28;10:2849. doi: 10.1038/s41467-019-10408-5

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — Role of UAF1 and RAD51AP1 in FANCD2 deubiquitination in cells. a HeLa cells depleted for endogenous UAF1 and stably expressing UAF1 (WT), UAF1^3A (3A), or UAF1^11A (11A) were examined for their level of ubiquitinated FANCD2 by western blotting. Ku86 was included as the loading control. b Survival of cells (from a) upon treatment with MMC or olaparib. The error bars represent mean values±S.D. of data from three independent experiments. c RAD51AP1 knockout HeLa cells depleted for endogenous UAF1 and stably expressing WT or mutant UAF1 with or without the co-expression of either RAD51AP1 (AP1) or the RAD51AP1^DM (DM) mutant were examined for their level of ubiquitinated FANCD2. Ku86 was included as the loading control. The error bars represent the mean+S.D. of data from three independent experiments. d Survival of cells (from c) upon treatment with MMC. The error bars represent mean values±S.D. of data from three independent experiments. e Model for the role of the USP1-UAF1-RAD51AP1 complex in the FA pathway. Mono-ubiquitination and deubiquitination of FANCD2 both occur on DNA. DNA binding by either UAF1 or RAD51AP1 is sufficient for targeting the trimeric DUB complex to DNA bound, mono-ubiquitinated FANCD2