Systematic analysis of barrier-forming FG hydrogels from Xenopus nuclear pore complexes

Nuclear pore complexes (NPCs) control the traffic between cell nucleus and cytoplasm. While facilitating translocation of nuclear transport receptors (NTRs) and NTR·cargo complexes, they suppress passive passage of macromolecules ⩾30 kDa. Previously, we reconstituted the NPC barrier as hydrogels com...

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Vydané v:	The EMBO journal Ročník 32; číslo 2; s. 204 - 218
Hlavní autori:	Labokha, Aksana A, Gradmann, Sabine, Frey, Steffen, Hülsmann, Bastian B, Urlaub, Henning, Baldus, Marc, Görlich, Dirk
Médium:	Journal Article
Jazyk:	English
Vydavateľské údaje:	Chichester, UK John Wiley & Sons, Ltd 23.01.2013 Nature Publishing Group UK Springer Nature B.V Nature Publishing Group
Predmet:	Active Transport, Cell Nucleus Amino Acid Sequence Animals beta Karyopherins - analysis beta Karyopherins - metabolism Cell Nucleus - chemistry Cell Nucleus - metabolism Cell Nucleus - ultrastructure Cells EMBO20 EMBO31 exportin FG hydrogel Glycine - chemistry Glycine - metabolism Hydrogels Hydrogels - analysis Hydrogels - chemistry Hydrogels - metabolism importin Membrane Microdomains - chemistry Membrane Microdomains - metabolism Membrane Microdomains - physiology Molecular Sequence Data Neurons Nuclear Pore - chemistry Nuclear Pore - metabolism Nuclear Pore - physiology nuclear pore complex Nuclear Pore Complex Proteins - analysis Nuclear Pore Complex Proteins - chemistry Nuclear Pore Complex Proteins - metabolism O-glycosylation Permeability Phenylalanine - chemistry Phenylalanine - metabolism Repetitive Sequences, Amino Acid Saccharomyces cerevisiae - chemistry Saccharomyces cerevisiae - metabolism Spectroscopy Translocation Xenopus - metabolism nuclear pore complex importin O‐glycosylation exportin FG hydrogel
ISSN:	0261-4189, 1460-2075, 1460-2075
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Popis
Shrnutí:	Nuclear pore complexes (NPCs) control the traffic between cell nucleus and cytoplasm. While facilitating translocation of nuclear transport receptors (NTRs) and NTR·cargo complexes, they suppress passive passage of macromolecules ⩾30 kDa. Previously, we reconstituted the NPC barrier as hydrogels comprising S. cerevisiae FG domains. We now studied FG domains from 10 Xenopus nucleoporins and found that all of them form hydrogels. Related domains with low FG motif density also substantially contribute to the NPC's hydrogel mass. We characterized all these hydrogels and observed the strictest sieving effect for the Nup98‐derived hydrogel. It fully blocks entry of GFP‐sized inert objects, permits facilitated entry of the small NTR NTF2, but arrests importin β‐type NTRs at its surface. O‐GlcNAc modification of the Nup98 FG domain prevented this arrest and allowed also large NTR·cargo complexes to enter. Solid‐state NMR spectroscopy revealed that the O‐GlcNAc‐modified Nup98 gel lacks amyloid‐like β‐structures that dominate the rigid regions in the S. cerevisiae Nsp1 FG hydrogel. This suggests that FG hydrogels can assemble through different structural principles and yet acquire the same NPC‐like permeability. The phenylalanine‐glycine (FG) domains of vertebrate nucleoporins assemble into hydrogels with different sieving characteristics for macromolecules. Nup98 forms the tightest filter, which is relieved by O‐linked glycosylation.
Bibliografia:	Supplementary InformationReview Process File ark:/67375/WNG-QB2M1DR8-L ArticleID:EMBJ2012302 istex:D91509F9D94954E7713FCAC9EB0B497026E26196 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 ObjectType-Article-2 ObjectType-Undefined-1 ObjectType-Feature-3 content type line 23
ISSN:	0261-4189 1460-2075 1460-2075
DOI:	10.1038/emboj.2012.302