Abstract
The Fanconi anemia (FA) pathway removes interstrand crosslinks (ICLs) between the Watson-Crick strands of the DNA double helix in humans. Central to the pathway is the FANCD2/FANCI complex, which must be loaded onto chromosomes. Here, we report the identification of a PP2A phosphatase complex, which specifically dephosphorylates an inhibitory cluster in FANCD2, thereby licensing its loading in response to DNA damage. We show that PP2A is required for normal monoubiquitination of the FANCD2/FANCI complex and for its loading onto chromosomes. We have fully reconstituted a coupled dephosphorylation-ubiquitination reaction in vitro using a highly purified PP2A complex. Using super-resolution live-cell single-molecule tracking, we show how PP2A switches on the FA pathway in response to ICLs and that cells are sensitive to ICL-forming drugs in the absence of PP2A. Our work uncovers a mechanism where PP2A facilitates the activation of the FA pathway by licensing chromosome loading of the FANCD2/FANCI complex.
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | 114971 |
| Tidsskrift | Cell Reports |
| Vol/bind | 43 |
| Udgave nummer | 11 |
| Antal sider | 22 |
| ISSN | 2639-1856 |
| DOI | |
| Status | Udgivet - 2024 |
Bibliografisk note
Funding Information:The authors would like to thank members of the Cohn laboratory for reading and discussing the manuscript. We are grateful to Stephan Uphoff for valuable advice on single-particle tracking. We are also grateful to staff at the Micron Imaging Facility for excellent help and support. We thank Luke Lavis (Janelia Research Campus) for providing the JF549 Halo ligand dye. This work was supported by grants UF100717 and UF150651 from the Royal Society (to M.A.C.), grants 153/092 and 0012197 from John Fell Oxford University Press Research Fund (to M.A.C.), grants OCRC 0213-MC and CRUKDF 0715-MC from the OCRC/CR-UK (to M.A.C.), a grant from the Medical Research Fund (to M.A.C.), grant MR/N021002/1 from the Medical Research Council (to M.A.C.), a Wellcome Trust Senior Research Fellowship , 210640/Z/18/Z (to M.A.C.), an MRC-Clarendon Scholarship (to D.L.-M.), a \u201CMutua Madrile\u00F1a\u201D Graduate Scholarship (to D.L.-M.), a fellowship from the Uehara Memorial Foundation (to A.J.-M.), and a Leathersellers\u2019 Company Scholarship via St. Catherine\u2019s College Oxford (to L.J.C.).
Publisher Copyright:
© 2024 The Author(s)