Assembly principle of a membrane-anchored nuclear pore basket scaffold

Nuclear pore complexes (NPCs) are membrane-embedded gatekeepers of traffic between the nucleus and cytoplasm. Key features of the NPC symmetric core have been elucidated, but little is known about the NPC basket, a prominent structure with numerous roles in gene expression. Studying the basket was hampered by its instability and connection to the inner nuclear membrane (INM). Here, we reveal the assembly principle of the yeast NPC basket by reconstituting a recombinant Nup60-Mlp1-Nup2 scaffold on a synthetic membrane. Nup60 serves as the basket’s flexible suspension cable, harboring an array of short linear motifs (SLiMs). These bind multivalently to the INM, the coiled-coil protein Mlp1, the FG-nucleoporin Nup2, and the NPC core. We suggest that SLiMs, embedded in disordered regions, allow the basket to adapt its structure in response to bulky cargo and changes in gene expression. Our study opens avenues for the higher-order reconstitution of basket-anchored NPC assemblies on membranes.

• Publication year

  2022

• Venue

  Science Advances

• Publication date

  2022-02-01

• Fields of study

  Biology, Medicine, Materials Science

• Identifiers
  DOI 10.1126/sciadv.abl6863PMID 35148185PMCID 8836807
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• The nuclear pore complex and the genome: organizing and regulatory principles.

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  2020influential reference
• Dissecting the Structural Dynamics of the Nuclear Pore Complex.

  2020cited by this paper
• Maturation Kinetics of a Multiprotein Complex Revealed by Metabolic Labeling.

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• Structure and Assembly of the Nuclear Pore Complex.

  2019cited by this paper
• Native Chromatin Proteomics Reveals a Role for Specific Nucleoporins in Heterochromatin Organization and Maintenance.

  2019cited by this paper
• In-depth and 3-Dimensional Exploration of the Budding Yeast Phosphoproteome

  2019influential reference
• The Structure of the Nuclear Pore Complex (An Update).

  2019cited by this paper
• ELM—the eukaryotic linear motif resource in 2020

  2019cited by this paper
• The Inner Nuclear Membrane Is a Metabolically Active Territory that Generates Nuclear Lipid Droplets

  2018influential reference
• The Dynamic Nature of the Nuclear Envelope.

  2018cited by this paper
• Integrative Structure and Functional Anatomy of a Nuclear Pore Complex

  2018cited by this paper
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  2017cited by this paper
• The roles of the nuclear pore complex in cellular dysfunction, aging and disease.

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  2017cited by this paper
• Mechanisms and functions of nuclear envelope remodelling

  2017cited by this paper
• The Nucleoporin Nup2 Contains a Meiotic-Autonomous Region that Promotes the Dynamic Chromosome Events of Meiosis

  2017cited by this paper
• Nuclear pore complexes and regulation of gene expression.

  2017cited by this paper
• Stoichiometry and compositional plasticity of the yeast nuclear pore complex revealed by quantitative fluorescence microscopy

  2017cited by this paper
• The Multiple Faces of Disordered Nucleoporins

  2016cited by this paper
• Architecture of the symmetric core of the nuclear pore

  2016cited by this paper
• Posttranslational marks control architectural and functional plasticity of the nuclear pore complex basket

  2016cited by this paper
• Transport Selectivity of Nuclear Pores, Phase Separation, and Membraneless Organelles.

  2016cited by this paper
• Nuclear pore assembly proceeds by an inside-out extrusion of the nuclear envelope

  2016cited by this paper
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• Linker Nups connect the nuclear pore complex inner ring with the outer ring and transport channel

  2015cited by this paper
• Nup153 Recruits the Nup107-160 Complex to the Inner Nuclear Membrane for Interphasic Nuclear Pore Complex Assembly.

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• In situ structural analysis of the human nuclear pore complex

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• Quantifying Nucleoporin Stoichiometry Inside Single Nuclear Pore Complexes In vivo

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• Short linear motifs: ubiquitous and functionally diverse protein interaction modules directing cell regulation.

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• Expression, purification and preliminary crystallographic analysis of the N-terminal domain of Trypanosoma brucei BILBO1.

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• A sub-nanometre view of how membrane curvature and composition modulate lipid packing and protein recruitment

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• The nuclear basket proteins Mlp1p and Mlp2p are part of a dynamic interactome including Esc1p and the proteasome

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• Nuclear Pore Scaffold Structure Analyzed by Super-Resolution Microscopy and Particle Averaging

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  2011cited by this paper
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  2010cited by this paper
• Cell cycle-dependent differences in nuclear pore complex assembly in metazoa.

  2010cited by this paper
• We thank S

  2010cited by this paper
• A general amphipathic α-helical motif for sensing membrane curvature

  2007cited by this paper
• Molecular mechanism of the nuclear protein import cycle

  2007cited by this paper
• A general amphipathic alpha-helical motif for sensing membrane curvature.

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• The solubility and stability of recombinant proteins are increased by their fusion to NusA.

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• Nuclear retention of unspliced mRNAs in yeast is mediated by perinuclear Mlp1.

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  2001cited by this paper
• The Yeast Nuclear Pore Complex: Composition, Architecture, and Transport Mechanism

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  2000cited by this paper
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• High-resolution field emission scanning electron microscope imaging of internal cell structures after Epon extraction from sections: a new approach to correlative ultrastructural and immunocytochemical studies.

  1993cited by this paper
• Toward a more complete 3-D structure of the nuclear pore complex.

  1991cited by this paper
• A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae.

  1989cited by this paper

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