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TitleThe DNA-damage kinase ATR activates the FANCD2-FANCI clamp by priming it for ubiquitination.
Journal, issue, pagesNat Struct Mol Biol, Vol. 29, Issue 9, Page 881-890, Year 2022
Publish dateSep 1, 2022
AuthorsTamara Sijacki / Pablo Alcón / Zhuo A Chen / Stephen H McLaughlin / Shabih Shakeel / Juri Rappsilber / Lori A Passmore /
PubMed AbstractDNA 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 ...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.
External linksNat Struct Mol Biol / PubMed:36050501 / PubMed Central
MethodsEM (single particle)
Resolution3.53 - 4.4 Å
Structure data

15101

8a2q

EMDB-15101, PDB-8a2q:
Structure of the DNA-bound FANCD2-FANCI complex containing phosphomimetic FANCI
Method: EM (single particle) / Resolution: 3.53 Å

EMDB-15102: FANCD2-FANCI complex containing FANCI (FANCI3D) that contains phosphomimetic mutations
Method: EM (single particle) / Resolution: 4.1 Å

EMDB-15103: ubiquitinated DNA-bound FANCD2-FANCI3D
Method: EM (single particle) / Resolution: 4.4 Å

Source
  • gallus gallus (chicken)
  • synthetic construct (others)
  • Homo sapiens (human)
KeywordsDNA BINDING PROTEIN / nucleic acid protein complex / DNA clamp / solenoid domains / Fanconi anemia / DNA repair / DNA damage / DNA inter strand crosslink / ubiquitination / ATR / pathway activation

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