Nucleocytoplasmic transport of intrinsically disordered proteins studied by high‐speed super‐resolution microscopy

Nucleoporins (Nups) represent a range of proteins most known for composing the macromolecular assembly of the nuclear pore complex (NPC). Among them, the family of intrinsically disordered proteins (IDPs) phenylalanine-glycine (FG) rich Nups, form the permeability barrier and coordinate the high-speed nucleocytoplasmic transport in a selective way. Those FG-Nups have been demonstrated to participate in various biological processes besides nucleocytoplasmic transport. The high number of accessible hydrophobic motifs of FG-Nups potentially gives rise to this multifunctionality, enabling them to form unique microenvironments. In this review, we discuss the multifunctionality of disordered and F-rich Nups and the diversity of their localizations, emphasizing the important roles of those Nups in various regulatory and metabolic processes.

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Simple biophysics underpins collective conformations of the intrinsically disordered proteins of the Nuclear Pore Complex

eLife ◽

10.7554/elife.10785 ◽

2016 ◽

Vol 5 ◽

Cited By ~ 44

Author(s):

Andrei Vovk ◽

Chad Gu ◽

Michael G Opferman ◽

Larisa E Kapinos ◽

Roderick YH Lim ◽

...

Keyword(s):

Spatial Organization ◽

Intrinsically Disordered Proteins ◽

Nucleocytoplasmic Transport ◽

Transport Proteins ◽

Disordered Proteins ◽

Biophysical Model ◽

Nuclear Pore ◽

Intrinsically Disordered

Nuclear Pore Complexes (NPCs) are key cellular transporter that control nucleocytoplasmic transport in eukaryotic cells, but its transport mechanism is still not understood. The centerpiece of NPC transport is the assembly of intrinsically disordered polypeptides, known as FG nucleoporins, lining its passageway. Their conformations and collective dynamics during transport are difficult to assess in vivo. In vitro investigations provide partially conflicting results, lending support to different models of transport, which invoke various conformational transitions of the FG nucleoporins induced by the cargo-carrying transport proteins. We show that the spatial organization of FG nucleoporin assemblies with the transport proteins can be understood within a first principles biophysical model with a minimal number of key physical variables, such as the average protein interaction strengths and spatial densities. These results address some of the outstanding controversies and suggest how molecularly divergent NPCs in different species can perform essentially the same function.

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2P094 Direct visualization of intrinsically disordered proteins PQBP-1 and FliK using high-speed atomic force microscopy(The 48th Annual Meeting of the Biophysical Society of Japan)

Seibutsu Butsuri ◽

10.2142/biophys.50.s98_4 ◽

2010 ◽

Vol 50 (supplement2) ◽

pp. S98

Author(s):

Noriyuki Kodera ◽

Tetsuya Mori ◽

Shino Mizuno ◽

Mineyuki Mizuguchi ◽

Shin-Ichi Aizawa ◽

...

Keyword(s):

Atomic Force Microscopy ◽

Annual Meeting ◽

High Speed ◽

Intrinsically Disordered Proteins ◽

Disordered Proteins ◽

Direct Visualization ◽

Force Microscopy ◽

Intrinsically Disordered ◽

Atomic Force ◽

Biophysical Society

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1H1522 High-speed AFM observation of intrinsically disordered proteins (NTAIL-GFP, PNT-GFP and SIC1-GFP)(Bioimaging,Oral Presentation,The 50th Annual Meeting of the Biophysical Society of Japan)

Seibutsu Butsuri ◽

10.2142/biophys.52.s34_2 ◽

2012 ◽

Vol 52 (supplement) ◽

pp. S34

Author(s):

Sujit Kumar Dora ◽

Noriyuki Kodera ◽

David Bloquel ◽

Johnny Habchi ◽

Antoine Gruet ◽

...

Keyword(s):

Annual Meeting ◽

Oral Presentation ◽

High Speed ◽

Intrinsically Disordered Proteins ◽

Disordered Proteins ◽

Intrinsically Disordered ◽

Biophysical Society ◽

Afm Observation

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1N1548 P79 Direct visualization of intrinsically disordered proteins PQBP-1, MeCP2, and FliK using high-speed atomic force microscopy(Bioimaging 1,The 49th Annual Meeting of the Biophysical Society of Japan)

Seibutsu Butsuri ◽

10.2142/biophys.51.s68_3 ◽

2011 ◽

Vol 51 (supplement) ◽

pp. S68

Author(s):

Tetsuya Mori ◽

Noriyuki Kodera ◽

Shino Mizuno ◽

Shin-Ichi Aizawa ◽

Mineyuki Mizuguchi ◽

...

Keyword(s):

Atomic Force Microscopy ◽

Annual Meeting ◽

High Speed ◽

Intrinsically Disordered Proteins ◽

Disordered Proteins ◽

Direct Visualization ◽

Force Microscopy ◽

Intrinsically Disordered ◽

Atomic Force ◽

Biophysical Society

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Interactions of nuclear transport factors and surface-conjugated FG nucleoporins: Insights and limitations

10.1101/435651 ◽

2018 ◽

Author(s):

Ryo Hayama ◽

Mirco Sorci ◽

John J. Keating ◽

Lee M. Hecht ◽

Joel L. Plawsky ◽

...

Keyword(s):

Protein Interactions ◽

Intrinsically Disordered Proteins ◽

Binding Mode ◽

Nucleocytoplasmic Transport ◽

Disordered Proteins ◽

Protein Protein Interactions ◽

Force Microscopy ◽

Intrinsically Disordered ◽

Mutant Variant

ABSTRACTProtein-protein interactions are central to biological processes and the methods to thoroughly characterize them are of great interest. In vitro methods to examine protein-protein interactions are generally categorized into two classes: in-solution and surface-based methods. Here, using the multivalent interactions involved in nucleocytoplasmic transport as a model system, we examined the utility of three surface-based methods in characterizing rapid interactions involving intrinsically disordered proteins: atomic force microscopy, quartz crystal microbalance with dissipation, and surface plasmon resonance. Although results were comparable to those of previous reports, the existence of previously overlooked mass transport limitations was revealed. Additional experiments with a loss-of-interaction mutant variant demonstrated the existence of additional physical events and an uncharacterized binding mode. These results suggest the binding events that take place on the surface are more complex than initially assumed, prompting a need for re-interpretation of previous data.

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