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BID-induced structural changes in BAK promote apoptosis

Nature Structural & Molecular Biology volume 20, pages 589–597 (2013)Cite this article

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Abstract

The BCL-2–family protein BAK is responsible for mitochondrial outer-membrane permeabilization (MOMP), which leads to apoptosis. The BCL-2 homology 3 (BH3)-only protein BID activates BAK to perform this function. We report the NMR solution structure of the human BID BH3–BAK complex, which identified the activation site at the canonical BH3-binding groove of BAK. Mutating the BAK BH1 in the groove prevented activation and MOMP but not the binding of BID. BAK BH3 mutations allowed BID binding and activation but blunted function by blocking BAK oligomerization. BAK activation follows a 'hit-and-run' mechanism whereby BID dissociates from the trigger site, which allows BAK oligomerization at an overlapping interface. In contrast, the BH3-only proteins NOXA and BAD are predicted to clash with the trigger site and are not activators of BAK. These findings provide insights into the early stages of BAK activation.

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Figure 1: BID BH3 binds the BC groove to directly activate BAK.
Figure 2: BID induces opening of the occluded BC groove in BAK.
Figure 3: Molecular determinants for BAK direct activation extend throughout BID BH3.
Figure 4: Activator–effector complexes support a hit-and-run activation mechanism.
Figure 5: Direct activation and oligomerization separated by point mutations in BAK.
Figure 6: NOXA and BAD are not direct activators of BAK.
Figure 7: Direct activation triggers BAK-mediated MOMP.

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Acknowledgements

We thank C. Dillon, L. McCormick and M. Yang for managing the mouse colony, R. Cassell and P. Rodriques of the St. Jude Hartwell Center for producing the peptides and R. Cross and G. Lennon of the St. Jude Flow Cytometry facility for cell sorting. This work was supported by US National Institutes of Health grants AI40646, GM52735 and GM096208 (D.R.G.) and R01CA082491 and 1R01GM083159 (R.W.K.) and by the American Lebanese Syrian Associated Charities.

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Authors and Affiliations

  1. Department of Immunology, St. Jude Children's Research Hospital, Memphis, Tennessee, USA

    Tudor Moldoveanu, Fabien Llambi, Patrick Fitzgerald & Douglas R Green

  2. Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, Tennessee, USA

    Christy R Grace & Richard W Kriwacki

  3. Hartwell Center for Bioinformatics and Biotechnology, St. Jude Children's Research Hospital, Memphis, Tennessee, USA

    Amanda Nourse

  4. Department of Biochemistry, McGill University, Montreal, Quebec, Canada

    Kalle Gehring

  5. Department of Microbiology, Immunology and Biochemistry, University of Tennessee Health Sciences Center, Memphis, Tennessee, USA

    Richard W Kriwacki

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Contributions

T.M., C.R.G., K.G. and R.W.K. contributed the NMR analyses, T.M., F.L. and P.F. the biochemical analyses and T.M. and A.N. the AUC analyses. All authors contributed conceptually to various aspects of the project. T.M., R.W.K. and D.R.G. wrote the manuscript.

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Correspondence to Douglas R Green.

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Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–6, Supplementary Tables 1–2 and Supplementary Note (PDF 1699 kb)

Supplementary Movie 1

Movie captures the conformational changes of apo to SAHBa–bound cBAK described in Fig. 2 andSupplementary Fig. 2. LSQMAN3 was used to generate the frames and Pymol to render them. (AVI 9300 kb)

Supplementary Movie 2

Movie captures the conformational changes of apo to SAHBa–bound cBAK from a side viewcompared to that of Supplementary Movie 1. (AVI 12207 kb)

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Moldoveanu, T., Grace, C., Llambi, F. et al. BID-induced structural changes in BAK promote apoptosis. Nat Struct Mol Biol 20, 589–597 (2013). https://doi.org/10.1038/nsmb.2563

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