Toward a more complete 3-D structure of the nuclear pore complex.
The nuclear pore complex (NPC) is a large supramolecular assembly embedded in the double-membraned nuclear envelope (NE) that plays a pivotal role in the exchange of macromolecules and particles between the nucleus and the cytoplasm. Applying various methods of sample preparation to Xenopus laevis whole nuclei and isolated NEs in combination with conventional transmission electron microscopy and digital image processing, we have characterized several distinct components of the NPC, including massive cytoplasmic and more tenuous nuclear rings, NPCs devoid of their cytoplasmic or both rings, and prominent "knobs" that protrude from the periphery of the NPC proper into the lumen of the NE. Moreover, by quick freezing/freeze drying/rotary metal shadowing isolated NEs, we have visualized two distinct types of NPC-associated filaments: (1) eight short, highly twisted filaments that project from the cytoplasmic ring and sometimes collapse into short cylinders; and (2) eight long, thin filaments that protrude from the nuclear ring and whose ends join to form a distal ring centered above the NPC such that the assembly resembles a "fishtrap." These nuclear fishtraps are sensitive to divalent cations: removal unfolds them and addition reforms them. The significance of these various structural components in terms of current NPC models is discussed, and the emerging asymmetry of the NPC relative to its nuclear and cytoplasmic face is stressed.
Jarnik
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M. E. Müller-Institute for High Resolution Electron Microscopy, University of Basel, Switzerland.
Aebi
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U
D000199:Actins; D000818:Animals; D015925:Cryopreservation; D003902:Detergents; D005612:Freeze Drying; D007091:Image Processing, Computer-Assisted; D008854:Microscopy, Electron; D009685:Nuclear Envelope / Q000502:physiology / Q000648:ultrastructure*; D012260:Ribonucleases; D013194:Staining and Labeling; D014982:Xenopus laevis
D000199:Actins; D003902:Detergents; D012260:Ribonucleases