Architecture of the nuclear pore complex coat
The nuclear pore complex (NPC) constitutes the sole gateway for bidirectional nucleocytoplasmic transport. Despite half a century of structural characterization, the architecture of the NPC remains unknown. Here, we present the crystal structure of a reconstituted ~400 kDa coat nucleoporin complex (CNC) from S. cerevisiae at a 7.4-Å resolution. The crystal structure revealed a curved Y-shaped architecture and the molecular details of the coat nucleoporin interactions forming the central "triskelion" of the Y. A structural comparison of the yeast CNC with an electron microscopy reconstruction of its human counterpart suggested the evolutionary conservation of the elucidated architecture. Moreover, 32 copies of the CNC crystal structure docked readily into a cryoelectron tomographic reconstruction of the fully-assembled human NPC, thereby accounting for ~16 MDa of its mass.
Additional Information© 2015 American Association for the Advancement of Science. Received for publication 18 September 2014. Accepted for publication 27 January 2015. Published Online February 12 2015. We thank C. J. Bley, W. M. Clemons, A. M. Davenport, O. Dreesen, A. Patke, D. C. Rees, S. O. Shan, P. Stravropoulos, and K. Thierbach for critical reading of the manuscript, K. Kato and K. Kato for their contributions at the initial stages of this project, L. N. Collins for technical support, D. King for mass spectrometry analysis, E. Hurt and P. Loppnau for material, S. Koide for providing the phage display library, S. Gräslund for the pSFV4 vector, P. Afonine for advice regarding structure refinement in PHENIX, and J. Kaiser and the scientific staff of SSRL Beamline 12-2 and the APS Beamline GM/CA-CAT for their support with x-ray diffraction measurements. We acknowledge the Gordon and Betty Moore Foundation, the Beckman Institute, and the Sanofi-Aventis Bioengineering Research Program for their support of the Molecular Observatory at the California Institute of Technology. The operations at the SSRL and APS are supported by the Department of Energy and the National Institutes of Health. T.S. was supported by a Postdoctoral Fellowship of the Deutsche Forschungsgemeinschaft. D.H.L. was supported by a National Institutes of Health Research Service Award (5 T32 GM07616). A.A.K. was supported by National Institutes of Health Awards (U01 GM094588, U54 GM087519) and the Chicago Biomedical Consortium. A.H. was supported by Caltech startup funds, the Albert Wyrick V Scholar Award of the V Foundation for Cancer Research, the 54th Mallinckrodt Scholar Award of the Edward Mallinckrodt, Jr. Foundation, and a Kimmel Scholar Award of the Sidney Kimmel Foundation for Cancer Research. The coordinates and structure factors have been deposited with the Protein Data Bank with accession codes 4XMM and 4XMN. The authors declare no financial conflicts of interest.
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