Integrated Structural Analysis of the Human Nuclear Pore Complex Scaffold

Abstract

The nuclear pore complex (NPC) is a fundamental component of all eukaryotic cells that facilitates nucleocytoplasmic exchange of macromolecules. It is assembled from multiple copies of about 30 nucleoporins. Due to its size and complex composition, determining the structure of the NPC is an enormous challenge, and the overall architecture of the NPC scaffold remains elusive. In this study, we have used an integrated approach based on electron tomography, single-particle electron microscopy, and crosslinking mass spectrometry to determine the structure of a major scaffold motif of the human NPC, the Nup107 subcomplex, in both isolation and integrated into the NPC. We show that 32 copies of the Nup107 subcomplex assemble into two reticulated rings, one each at the cytoplasmic and nuclear face of the NPC. This arrangement may explain how changes of the diameter are realized that would accommodate transport of huge cargoes.

Integrative structure and functional anatomy of a nuclear pore complex

“…In yeast, each outer ring is formed by 8 copies of the Nup84 complex (Figs. 3F, 4), whereas in vertebrates each outer ring contains 16 copies of the equivalent Y-shaped complex arranged in two interlocked rings 17,40 . Moreover, we see neither an additional copy of Nup157/Nup170 connecting the outer and inner rings nor Nup188/Nup192 in the outer rings, as indicated in humans 6 (Fig.…”

Architecture of the yeast Elongator complex

“…The fits were then clustered, in the case of Elp123 sub‐complex taking the twofold symmetry axis into account. The P ‐values for the cross‐correlation scores were calculated by transforming to z ‐scores (Fisher's z ‐transform) and centering, and fitting an empirical null distribution from which two‐sided P ‐values were computed 64.…”

Molecular interactions of theSaccharomyces cerevisiaeAtg1 complex provide insights into assembly and regulatory mechanisms

“…CXMS has emerged as an important structural technique to identify potential subunit interaction interfaces in the context of the intact, fully assembled macromolecular assembly. [41][42][43] Using this novel approach, we were able to comprehensively map the subunit interaction interfaces of a reconstituted minimal Atg1 pentameric complex from the model organism S. cerevisiae (Fig. 7).…”