Siah ubiquitin ligase is structurally related to TRAF and modulates TNF-α signaling

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Abstract

Members of the Siah (seven in absentia homolog) family of RING domain proteins are components of E3 ubiquitin ligase complexes that catalyze ubiquitination of proteins. We have determined the crystal structure of the substrate-binding domain (SBD) of murine Siah1a to 2.6 Å resolution. The structure reveals that Siah is a dimeric protein and that the SBD adopts an eight-stranded β-sandwich fold that is highly similar to the TRAF-C region of TRAF (TNF-receptor associated factor) proteins. The TRAF-C region interacts with TNF-α receptors and TNF-receptor associated death-domain (TRADD) proteins; however, our findings indicate that these interactions are unlikely to be mimicked by Siah. The Siah structure also reveals two novel zinc fingers in a region with sequence similarity to TRAF. We find that the Siah1a SBD potentiates TNF-α-mediated NF-κB activation. Therefore, Siah proteins share important similarities with the TRAF family of proteins, including their overall domain architecture, three-dimensional structure and functional activity.

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Figure 1: Primary structure of Siah.
Figure 2: Siah1a SBD forms a dimer in solution.
Figure 3: Structure of Siah1a SBD.
Figure 4: Similarities between Siah and TRAF.
Figure 5: Surface features of Siah SBD.

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Acknowledgements

We thank K. Wilson, F. Katsis and B. Cromer (St. Vincent's Institute of Medical Research) for assistance with mass spectrometry and protein expression, and M. Pardee for N-terminal sequencing. We thank H. Tong and other staff at BioCARS for their help with data collection during our visit to the Advanced Photon Source. Access to BioCARS Sector 14 at the Advanced Photon Source at Argonne, Illinois, was provided by the Australian Synchrotron Research Program, which is funded by the Commonwealth of Australia as a Major National Research Facility. BioCARS Sector 14 is supported by the U.S. National Institutes of Health, National Center for Research Resources. The Advanced Photon Source is supported by the U.S. Department of Energy, Basic Energy Sciences, Office of Energy Research. M.W.P. is an Australian Research Council Senior Research Fellow, D.D.L.B. is a National Health and Medical Research Council of Australia Senior Research Fellow and D.S. is an American Heart Association Kenner Fellow. This work was additionally supported by grants from the NIH (NCI) (D.S.).

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Correspondence to David D.L. Bowtell.

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Polekhina, G., House, C., Traficante, N. et al. Siah ubiquitin ligase is structurally related to TRAF and modulates TNF-α signaling. Nat Struct Mol Biol 9, 68–75 (2002) doi:10.1038/nsb743

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