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Molecular mechanism of Reaper-Grim-Hid-mediated suppression of DIAP1-dependent Dronc ubiquitination

Nature Structural & Molecular Biology volume 10, pages 892–898 (2003)Cite this article

Abstract

The inhibitor of apoptosis protein DIAP1 inhibits Dronc-dependent cell death by ubiquitinating Dronc. The pro-death proteins Reaper, Hid and Grim (RHG) promote apoptosis by antagonizing DIAP1 function. Here we report the structural basis of Dronc recognition by DIAP1 as well as a novel mechanism by which the RHG proteins remove DIAP1-mediated downregulation of Dronc. Biochemical and structural analyses revealed that the second BIR (BIR2) domain of DIAP1 recognizes a 12-residue sequence in Dronc. This recognition is essential for DIAP1 binding to Dronc, and for targeting Dronc for ubiquitination. Notably, the Dronc-binding surface on BIR2 coincides with that required for binding to the N termini of the RHG proteins, which competitively eliminate DIAP1-mediated ubiquitination of Dronc. These observations reveal the molecular mechanisms of how DIAP1 recognizes Dronc, and more importantly, how the RHG proteins remove DIAP1-mediated ubiquitination of Dronc.

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Figure 1: The recognition of Dronc by DIAP1 is restricted to the second BIR domain (BIR2) of DIAP1 and a 12-residue peptide in the linker sequence between the CARD domain and the catalytic subunit of Dronc.
Figure 2: Recognition of Dronc is necessary for DIAP1-mediated negative regulation of Dronc in vivo.
Figure 3: Structural mechanism of Dronc recognition by DIAP1.
Figure 4: Dronc and Hid compete with each other for binding to the same surface groove on the BIR2 domain of DIAP1.
Figure 5: Molecular mechanism of the removal of DIAP1-mediated Dronc ubiquitination by the pro-apoptosis protein Hid.
Figure 6: A schematic representation of the regulation of Dronc by DIAP1 and the RHG proteins.

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Acknowledgements

We thank L. Walsh and others at CHESS for help with beam time and data collection. This research was supported by US National Institutes of Health grants.

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  1. Jijie Chai and Nieng Yan: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Biology, Princeton University, Lewis Thomas Laboratory, Washington Road, Princeton, 08544, New Jersey, USA

    Jijie Chai, Nieng Yan, Wenyu Li & Yigong Shi

  2. Division of Biology, MC 156-29, California Institute of Technology, Pasadena, 91125, California, USA

    Jun R Huh & Bruce A Hay

  3. Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing, 100084, China

    Jia-Wei Wu

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Correspondence to Yigong Shi.

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Chai, J., Yan, N., Huh, J. et al. Molecular mechanism of Reaper-Grim-Hid-mediated suppression of DIAP1-dependent Dronc ubiquitination. Nat Struct Mol Biol 10, 892–898 (2003). https://doi.org/10.1038/nsb989

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