Size-dependent leak of soluble and membrane proteins through the yeast nuclear pore complex

• Published in: Molecular biology of the cell 2015-04, Vol.26 (7), p.1386-1394
• Main Authors: Popken, Petra, Ghavami, Ali, Onck, Patrick R, Poolman, Bert, Veenhoff, Liesbeth M
• Format: Article
• Language: English
• Subjects: Diffusion; Membrane Proteins - metabolism; Microscopy, Fluorescence; Molecular Dynamics Simulation; Nuclear Pore - metabolism; Protein Transport; Saccharomyces cerevisiae - metabolism
• ISSN: 1059-1524; 1939-4586
• DOI: 10.1091/mbc.e14-07-1175

Nuclear pore complexes (NPCs) allow selective import and export while forming a barrier for untargeted proteins. Using fluorescence microscopy, we measured in vivo the permeability of the Saccharomyces cerevisiae NPC for multidomain proteins of different sizes and found that soluble proteins of 150 kDa and membrane proteins with an extralumenal domain of 90 kDa were still partly localized in the nucleus on a time scale of hours. The NPCs thus form only a weak barrier for the majority of yeast proteins, given their monomeric size. Using FGΔ-mutant strains, we showed that specific combinations of Nups, especially with Nup100, but not the total mass of FG-nups per pore, were important for forming the barrier. Models of the disordered phase of wild-type and mutant NPCs were generated using a one bead per amino acid molecular dynamics model. The permeability measurements correlated with the density predictions from coarse-grained molecular dynamics simulations in the center of the NPC. The combined in vivo and computational approach provides a framework for elucidating the structural and functional properties of the permeability barrier of nuclear pore complexes.