Abstract
The cullin 4–DNA-damage-binding protein 1 (CUL4–DDB1) ubiquitin ligase machinery regulates diverse cellular functions and can be subverted by pathogenic viruses. Here we report the crystal structure of DDB1 in complex with a central fragment of hepatitis B virus X protein (HBx), whose DDB1-binding activity is important for viral infection. The structure reveals that HBx binds DDB1 through an α-helical motif, which is also found in the unrelated paramyxovirus SV5-V protein despite their sequence divergence. Our structure-based functional analysis suggests that, like SV5-V, HBx captures DDB1 to redirect the ubiquitin ligase activity of the CUL4–DDB1 E3 ligase. We also identify the α-helical motif shared by these viral proteins in the cellular substrate–recruiting subunits of the E3 complex, the DDB1–CUL4-associated factors (DCAFs) that are functionally mimicked by the viral hijackers. Together, our studies reveal a common yet promiscuous structural element that is important for the assembly of cellular and virally hijacked CUL4–DDB1 E3 complexes.
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Acknowledgements
We thank Advanced Light Source synchrotron beamline staff for assistance with data collection; all members of the Zheng laboratory for invaluable discussions; W. Xu for help and support in our research; and O. Leupin (Novartis) for the siRNA-resistant DDB1 clone. N.Z. is a Pew scholar. This study is supported by the Howard Hughes Medical Institute and by a Burroughs Wellcome Fund Investigators in the Pathogenesis of Infectious Disease award and US National Institutes of Health grant CA107134 to N.Z., and Swiss National Science Foundation grants 3100A0-112496 and 31003A-127384 to M.S.
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T.L. designed and performed crystallographic studies, sequence analyses and GST pulldown assays. M.S., E.I.R. and P.C.v.B. designed experiments for the functional studies of the viral proteins and analyzed the data. E.I.R. and P.C.v.B. carried out the experiments. N.Z. and M.S. supervised the project and are the principal manuscript authors.
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Li, T., Robert, E., van Breugel, P. et al. A promiscuous α-helical motif anchors viral hijackers and substrate receptors to the CUL4–DDB1 ubiquitin ligase machinery. Nat Struct Mol Biol 17, 105–111 (2010). https://doi.org/10.1038/nsmb.1719
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DOI: https://doi.org/10.1038/nsmb.1719
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