Abstract
The Fanconi anemia (FA) pathway is activated in response to DNA damage, leading to monoubiquitination of the substrates FANCI and FANCD2 by the FA core complex. Here we report the crystal structure of FANCL, the catalytic subunit of the FA core complex, at 3.2 Å. The structure reveals an architecture fundamentally different from previous sequence-based predictions. The molecule is composed of an N-terminal E2-like fold, which we term the ELF domain, a novel double-RWD (DRWD) domain, and a C-terminal really interesting new gene (RING) domain predicted to facilitate E2 binding. Binding assays show that the DRWD domain, but not the ELF domain, is responsible for substrate binding.
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Acknowledgements
We thank M. Way and F. Pinto for help with improving the manuscript, V. Chaugule for critical comments, discussion and technical assistance, L. Wood for assistance with sequence analysis and S. Kjaer and S. Kisakye-Nambozo of the Protein Production Facility for generation of the baculoviruses and subsequent Sf9 infection. All authors are funded by Cancer Research UK.
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A.R.C. crystallized, solved, refined and analyzed the FANCL structure and performed the biochemical experiments. All authors cloned, expressed and purified proteins. H.W. designed and supervised the study. A.R.C. and H.W. wrote the manuscript.
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Cole, A., Lewis, L. & Walden, H. The structure of the catalytic subunit FANCL of the Fanconi anemia core complex. Nat Struct Mol Biol 17, 294–298 (2010). https://doi.org/10.1038/nsmb.1759
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DOI: https://doi.org/10.1038/nsmb.1759
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