TY - JOUR
T1 - The DNA-damage kinase ATR activates the FANCD2-FANCI clamp by priming it for ubiquitination
AU - Sijacki, Tamara
AU - Alcón, Pablo
AU - Chen, Zhuo A.
AU - Mclaughlin, Stephen H.
AU - Shakeel, Shabih
AU - Rappsilber, Juri
AU - Passmore, Lori A.
N1 - Funding Information:
We are grateful to C. Johnson, K. Nguyen, R. Williams, K. J. Patel and members of the Passmore laboratory for assistance and advice; the MRC-LMB EM facility for access and support with EM sample preparation and data collection; J. Rodriguez Molina and V. Chandrasekaran (MRC-LMB) for assisting with cryo-EM data collection; J. Grimmett and T. Darling (MRC-LMB scientific computation); and J.G. Shi (MRC-LMB baculovirus) for support. We acknowledge Diamond Light Source for access to eBIC (proposal BI23268) funded by the Wellcome Trust, MRC and Biotechnology and Biological Sciences Research Council. This work was supported by the MRC as part of UK Research and Innovation, MRC file reference number MC_U105192715 (L.A.P.); a PhD studentship from the Cambridge Trust (T.S.); an EMBO Long-Term Fellowship, grant ALTF 692-2018 (P.A.); and the Deutsche Forschungsgemeinschaft (German Research Foundation) grant no. 329673113 (J.R.). The Wellcome Centre for Cell Biology is supported by core funding from the Wellcome Trust, grant no. 203149 (J.R.).
Publisher Copyright:
© 2022, The Author(s), under exclusive licence to Springer Nature America, Inc.
PY - 2022/9/1
Y1 - 2022/9/1
N2 - DNA interstrand cross-links are tumor-inducing lesions that block DNA replication and transcription. When cross-links are detected at stalled replication forks, ATR kinase phosphorylates FANCI, which stimulates monoubiquitination of the FANCD2–FANCI clamp by the Fanconi anemia core complex. Monoubiquitinated FANCD2–FANCI is locked onto DNA and recruits nucleases that mediate DNA repair. However, it remains unclear how phosphorylation activates this pathway. Here, we report structures of FANCD2–FANCI complexes containing phosphomimetic FANCI. We observe that, unlike wild-type FANCD2–FANCI, the phosphomimetic complex closes around DNA, independent of the Fanconi anemia core complex. The phosphomimetic mutations do not substantially alter DNA binding but instead destabilize the open state of FANCD2–FANCI and alter its conformational dynamics. Overall, our results demonstrate that phosphorylation primes the FANCD2–FANCI clamp for ubiquitination, showing how multiple posttranslational modifications are coordinated to control DNA repair.
AB - DNA interstrand cross-links are tumor-inducing lesions that block DNA replication and transcription. When cross-links are detected at stalled replication forks, ATR kinase phosphorylates FANCI, which stimulates monoubiquitination of the FANCD2–FANCI clamp by the Fanconi anemia core complex. Monoubiquitinated FANCD2–FANCI is locked onto DNA and recruits nucleases that mediate DNA repair. However, it remains unclear how phosphorylation activates this pathway. Here, we report structures of FANCD2–FANCI complexes containing phosphomimetic FANCI. We observe that, unlike wild-type FANCD2–FANCI, the phosphomimetic complex closes around DNA, independent of the Fanconi anemia core complex. The phosphomimetic mutations do not substantially alter DNA binding but instead destabilize the open state of FANCD2–FANCI and alter its conformational dynamics. Overall, our results demonstrate that phosphorylation primes the FANCD2–FANCI clamp for ubiquitination, showing how multiple posttranslational modifications are coordinated to control DNA repair.
U2 - 10.1038/s41594-022-00820-9
DO - 10.1038/s41594-022-00820-9
M3 - Article
C2 - 36050501
VL - 29
SP - 881
EP - 890
JO - Nature Structural & Molecular Biology
JF - Nature Structural & Molecular Biology
SN - 1545-9993
IS - 9
ER -