Abstract
The execution of apoptosis or programmed cell death comprises both caspase-dependent and caspase-independent processes. Apoptosis inducing factor (AIF) was identified as a major player in caspase-independent cell death. It induces chromatin condensation and initial DNA cleavage via an unknown molecular mechanism. Here we report the crystal structure of human AIF at 1.8 Å resolution. The structure reveals the presence of a strong positive electrostatic potential at the AIF surface, although the calculated isoelectric point for the entire protein is neutral. We show that recombinant AIF interacts with DNA in a sequence-independent manner. In addition, in cells treated with an apoptotic stimulus, endogenous AIF becomes co-localized with DNA at an early stage of nuclear morphological changes. Structure-based mutagenesis shows that DNA-binding defective mutants of AIF fail to induce cell death while retaining nuclear translocation. The potential DNA-binding site identified from mutagenesis also coincides with computational docking of a DNA duplex. These observations suggest that AIF-induced nuclear apoptosis requires a direct interaction with DNA.
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Acknowledgements
We thank N. Lue and J. Wang for discussions; the laboratories of H. Robertson, J. Darnel, S. Chen-Kiang and W. Muller for technical help and staff at the advanced photon source for assistance with data collection. This work was partially supported by a special grant from the Ligue contre le Cancer and the European Commission. H.Y. is a Revson postdoctoral fellow and H.W. is a Pew scholar of biomedical sciences and a Rita Allen Scholar.
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Ye, H., Cande, C., Stephanou, N. et al. DNA binding is required for the apoptogenic action of apoptosis inducing factor. Nat Struct Mol Biol 9, 680–684 (2002). https://doi.org/10.1038/nsb836
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DOI: https://doi.org/10.1038/nsb836
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