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Related Concept Videos

Nuclear Protein Sorting01:34

Nuclear Protein Sorting

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Nuclear protein sorting is the selective trafficking of histones, polymerases, gene regulatory proteins into the nucleus and exporting RNAs and ribosomes to the cytosol. It is a tightly controlled process that regulates gene expression within a cell.
Proteins targeted to the nucleus carry nuclear localization signals or NLS recognized by import receptors in the cytosol. Similarly, proteins with nuclear export signals are recognized by export receptors. Import and export receptors are...
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Before mRNAs are exported to the cytoplasm, it is crucial to check each mRNA for structural and functional integrity. Eukaryotic cells use several different mechanisms, collectively known as mRNA surveillance, to look for irregularities in mRNAs. Irregular or aberrant mRNA are rapidly degraded by various enzymes. If a defective mRNA escapes the surveillance, it would be translated into a protein which would either be non-functional or not function properly. One of the primary irregularities in...
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Nuclear protein sorting regulates nucleus composition and gene expression, crucial for determining the fate of a eukaryotic cell. Hence, the entry and exit of molecules across the nuclear envelope is a tightly controlled process. Nuclear protein sorting can be inhibited by one of the following ways: 1) masking cargo signal sequences, 2) modifying the nuclear receptor's affinity for cargo, 3) controlling the nuclear pore size, 4) retaining the cargo during its transit to the cytosol or the...
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The eukaryotic nucleus is a double membrane-bound organelle that contains nearly all of the cell’s genetic material in the form of chromosomes. It is rightly called the “brain” of the cell as it shoulders the responsibility of responding to various physiological processes, stress, altered metabolic conditions, and other cellular signals. 
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The nucleus is a membrane-bound organelle that acts as a control center in a eukaryotic cell. It contains chromosomal DNA, which controls gene expression and precisely regulates the production of proteins within the cell. In contrast, the DNA inside the mitochondria and chloroplast only carries out functions that are specific to those organelles.
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Related Experiment Video

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Single-Molecule Imaging of Nuclear Transport
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Single-Molecule Imaging of Nuclear Transport

Published on: June 9, 2010

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Imaging the assembly, structure, and function of the nuclear pore inside cells.

Shotaro Otsuka1, Anna Szymborska1, Jan Ellenberg1

  • 1Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.

Methods in Cell Biology
|May 27, 2014
PubMed
Summary

We developed a new method using multicolor 4D imaging to analyze nuclear pore complex (NPC) assembly kinetics in live cells. This technique confirms conserved NPC assembly across different mammalian cells after cell division.

Keywords:
Live-cell imagingNuclear envelopeNuclear transportNucleoporinParticle averagingPhotoswitchingSTORMSuper-resolution microscopy

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biophysics

Background:

  • The nuclear pore complex (NPC) regulates transport between the nucleus and cytoplasm.
  • NPCs and the nuclear envelope (NE) disassemble and reassemble during cell division in higher eukaryotes.
  • Live-cell imaging is essential for studying these dynamic cellular processes.

Purpose of the Study:

  • To present a novel method for kinetic analysis of postmitotic NPC assembly.
  • To assess the reestablishment of nuclear transport competence after cell division.
  • To demonstrate the conservation of NPC assembly mechanisms in different mammalian cell types.

Main Methods:

  • Multicolor 4D imaging combined with photoswitching for live-cell analysis.
  • Kinetic analysis of nuclear pore complex assembly and transport function.
  • Application of previously established methods to HeLa cells.
  • Detailed presentation of stochastic super-resolution microscopy combined with single-particle averaging for NPC molecular organization studies.

Main Results:

  • The study presents a detailed method for analyzing the kinetics of nuclear pore complex assembly.
  • The reestablishment of nuclear transport competence was analyzed in intact cells.
  • NPC assembly was demonstrated to be conserved across different mammalian cell types (rat kidney and HeLa cells).
  • The molecular organization of the NPC was analyzed using super-resolution microscopy and single-particle averaging.

Conclusions:

  • The developed multicolor 4D imaging and photoswitching method allows for kinetic analysis of NPC assembly in live cells.
  • NPC assembly and the restoration of nuclear transport are conserved processes in mammalian cells.
  • Advanced microscopy techniques provide detailed insights into the molecular organization of the NPC.