eLife | |
Monoubiquitination by the human Fanconi anemia core complex clamps FANCI:FANCD2 on DNA in filamentous arrays | |
Andrew Leis1  Michael W Parker2  Sylvie van Twest3  Vincent J Murphy3  Michael Sharp3  Rohan Bythell-Douglas3  Andrew J Deans4  Wayne Crismani4  Winnie Tan4  | |
[1] Bio21 Institute, University of Melbourne, Parkville, Australia;Bio21 Institute, University of Melbourne, Parkville, Australia;Structural Biology Unit, St. Vincent’s Institute of Medical Research, Fitzroy, Australia;Genome Stability Unit, St. Vincent’s Institute of Medical Research, Fitzroy, Australia;Genome Stability Unit, St. Vincent’s Institute of Medical Research, Fitzroy, Australia;Department of Medicine (St. Vincent’s Health), The University of Melbourne, Melbourne, Australia; | |
关键词: Fanconi anemia; DNA repair; biochemistry; ubiquitin; Human; Xenopus; | |
DOI : 10.7554/eLife.54128 | |
来源: publisher | |
【 摘 要 】
FANCI:FANCD2 monoubiquitination is a critical event for replication fork stabilization by the Fanconi anemia (FA) DNA repair pathway. It has been proposed that at stalled replication forks, monoubiquitinated-FANCD2 serves to recruit DNA repair proteins that contain ubiquitin-binding motifs. Here, we have reconstituted the FA pathway in vitro to study functional consequences of FANCI:FANCD2 monoubiquitination. We report that monoubiquitination does not promote any specific exogenous protein:protein interactions, but instead stabilizes FANCI:FANCD2 heterodimers on dsDNA. This clamping requires monoubiquitination of only the FANCD2 subunit. We further show using electron microscopy that purified monoubiquitinated FANCI:FANCD2 forms filament-like arrays on long dsDNA. Our results reveal how monoubiquitinated FANCI:FANCD2, defective in many cancer types and all cases of FA, is activated upon DNA binding.
【 授权许可】
CC BY
【 预 览 】
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RO202004214226836ZK.pdf | 5046KB | download |