scholarly journals Super-resolved 3D Tracking of Cargo Transport Through Nuclear Pore Complexes by Astigmatism Imaging

2022 ◽  
Author(s):  
Rajdeep Chowdhury ◽  
Abhishek Sau ◽  
Siegfried M. Musser
Keyword(s):  
Adaptive Optics ◽  
3D Visualization ◽  
Nuclear Pore ◽  
Nuclear Pore Complexes ◽  
3D Tracking ◽  
Permeabilized Cells ◽  
Double Ring ◽  
Cargo Transport ◽  
Pore Complexes ◽  
20 Nm

Abstract This protocol describes a two-color astigmatic imaging approach that enables direct 3D visualization of cargo transport trajectories relative to a super-resolved octagonal double-ring scaffold structure of the nuclear pore complex (NPC). Though astigmatism imaging is commonly achieved via a cylindrical lens, this protocol utilizes an adaptive optics (AO) system, which enables optimization of the astigmatism for the precision needs of the experiment as well as correction of the focal mismatch arising from chromatic aberrations in multi-color applications. With this approach, single particle spatial precision values in x, y, and z are typically 5-20 nm, and these depend on astigmatism, photon level and position in z. The method enables resolution of transport conduits through the ~60 nm diameter pore of NPCs by particle tracking on the millisecond timescale. The success of this approach is enabled by the high rigidity of fully active NPCs within the nuclear envelope of permeabilized cells. For a detailed application of this protocol, please refer to https://www.nature.com/articles/s41556-021-00815-6. The figure and table numbers in this protocol that are indicated with an “NCB” prefix (e.g., NCB Figure X) refer to the figures and table in this reference paper.

scholarly journals Recapitulation of selective nuclear import and export with a perfectly repeated 12mer GLFG peptide

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Sheung Chun Ng ◽  
Thomas Güttler ◽  
Dirk Görlich
Keyword(s):  
Nucleocytoplasmic Transport ◽  
Nuclear Pore ◽  
Permeability Barrier ◽  
Nuclear Pore Complexes ◽  
Cell Nuclei ◽  
Cargo Transport ◽  
Fast Transport ◽  
Import And Export ◽  
Pore Complexes ◽  
Fundamental Requirement

AbstractThe permeability barrier of nuclear pore complexes (NPCs) controls nucleocytoplasmic transport. It retains inert macromolecules while allowing facilitated passage of importins and exportins, which in turn shuttle cargo into or out of cell nuclei. The barrier can be described as a condensed phase assembled from cohesive FG repeat domains. NPCs contain several distinct FG domains, each comprising variable repeats. Nevertheless, we now found that sequence heterogeneity is no fundamental requirement for barrier function. Instead, we succeeded in engineering a perfectly repeated 12mer GLFG peptide that self-assembles into a barrier of exquisite transport selectivity and fast transport kinetics. This barrier recapitulates RanGTPase-controlled importin- and exportin-mediated cargo transport and thus represents an ultimately simplified experimental model system. An alternative proline-free sequence forms an amyloid FG phase. Finally, we discovered that FG phases stain bright with ‘DNA-specific’ DAPI/ Hoechst probes, and that such dyes allow for a photo-induced block of nuclear transport.

scholarly journals Nucleocytoplasmic Communication in Healthy and Diseased Plant Tissues

2021 ◽  
Vol 12 ◽  
Author(s):  
Daniel Lüdke ◽  
Philipp F. W. Rohmann ◽  
Marcel Wiermer
Keyword(s):  
Nuclear Pore ◽  
Diseased Plant ◽  
Nuclear Pore Complexes ◽  
Cargo Transport ◽  
Independent Functions ◽  
Transport Receptors ◽  
Associated Proteins ◽  
Pore Complexes ◽  
Bidirectional Exchange ◽  
Cytoplasmic Processes

The double membrane of the nuclear envelope (NE) constitutes a selective compartment barrier that separates nuclear from cytoplasmic processes. Plant viability and responses to a changing environment depend on the spatial communication between both compartments. This communication is based on the bidirectional exchange of proteins and RNAs and is regulated by a sophisticated transport machinery. Macromolecular traffic across the NE depends on nuclear transport receptors (NTRs) that mediate nuclear import (i.e. importins) or export (i.e. exportins), as well as on nuclear pore complexes (NPCs) that are composed of nucleoporin proteins (NUPs) and span the NE. In this review, we provide an overview of plant NPC- and NTR-directed cargo transport and we consider transport independent functions of NPCs and NE-associated proteins in regulating plant developmental processes and responses to environmental stresses.

Nuclear Envelope Dynamics During Mitosis

1988 ◽  
Vol 46 ◽  
pp. 224-225
Author(s):  
Brian Burke
Keyword(s):  
Nuclear Envelope ◽  
Membrane Vesicles ◽  
Nuclear Pore ◽  
Nuclear Pore Complexes ◽  
Animal Cells ◽  
Interphase Chromosome ◽  
Lamin B ◽  
Chromosome Architecture ◽  
Complex Membrane ◽  
Pore Complexes

The nuclear envelope is a complex membrane structure that forms the boundary of the nuclear compartment in eukaryotes. It regulates the passage of macromolecules between the two compartments and may be important for organizing interphase chromosome architecture. In interphase animal cells it forms a remarkably stable structure consisting of a double membrane ouerlying a protein meshwork or lamina and penetrated by nuclear pore complexes. The latter form the channels for nucleocytoplasmic exchange of macromolecules, At the onset of mitosis, however, it rapidly disassembles, the membranes fragment to yield small vesicles and the lamina, which is composed of predominantly three polypeptides, lamins R, B and C (MW approx. 74, 68 and 65 kDa respectiuely), breaks down. Lamins B and C are dispersed as monomers throughout the mitotic cytoplasm, while lamin B remains associated with the nuclear membrane vesicles.

Pore relations: nuclear pore complexes and nucleocytoplasmic exchange

2000 ◽  
Vol 36 ◽  
pp. 75-88 ◽  
Author(s):  
Michael P. Rout ◽  
John D. Aitchison
Keyword(s):  
Nuclear Pore ◽  
Nuclear Pore Complexes ◽  
Pore Complexes

Quantifying tagged mRNA export flux via nuclear pore complexes in single live cells

2021 ◽  
Vol 545 ◽  
pp. 138-144
Author(s):  
Yueyue Jing ◽  
Yilin Lv ◽  
Jingya Ye ◽  
Longfang Yao ◽  
Liwen Chen ◽  
...  
Keyword(s):  
Mrna Export ◽  
Live Cells ◽  
Nuclear Pore ◽  
Nuclear Pore Complexes ◽  
Export Flux ◽  
Pore Complexes
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