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Related Concept Videos

Nuclear Localization Signals and Import01:46

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Proteins targeted to the nucleus carry short stretches of amino acid sequences called the nuclear localization signal or NLS. Classical nuclear localization signals are of two types: monopartite and bipartite NLS. Monopartite classical NLS (cNLS) consists of a single cluster of 4-8 amino acids. Bipartite cNLS consists of two clusters of  2-3 amino acids and a 9-12 residue long proline-rich linker bridging the two clusters. Signal clusters are rich in positively charged amino acids such as...
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The nucleolus is the most prominent substructure of the nucleus. When it was first discovered, it was considered to be an isolated organelle that forms fibrils and granules. In 1931, the relationship between the nucleolus and chromosomes was first described by Heitz. He observed that the appearance and size of nucleolus varies depending on the stage of the cell cycle. He also noticed constricted regions on different chromosomes clustered together at definite cell cycle stages. These regions,...
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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
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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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Updated: Apr 25, 2026

Using LEXY and LINuS Optogenetics Tools and Automated Image Analysis to Quantify Nucleocytoplasmic Transport Dynamics in Live Cells
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Using LEXY and LINuS Optogenetics Tools and Automated Image Analysis to Quantify Nucleocytoplasmic Transport Dynamics in Live Cells

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A Toolbox for Quantifying Nuclear and Nucleolar Protein Accumulation Using NLS and NoLS Fusion Reporters.

Eugene A Arifulin1, Yana R Musinova1, Ulyana V Matveeva2

  • 1Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, Russia.

Traffic (Copenhagen, Denmark)
|April 24, 2026
PubMed
Summary
This summary is machine-generated.

Researchers developed a standardized protocol using fluorescent protein fusions to measure nuclear and nucleolar accumulation. This method enhances the reliable quantification of nuclear transport mechanisms and signal sequence function in vivo.

Keywords:
cytoplasmimportinslive‐cell imagingnuclear localization signal (NLS)nucleolar localization signal (NoLS)nucleolusnucleus

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Fluorescent proteins fused to nuclear localization signals (NLSs) or nucleolar localization signals (NoLSs) are vital tools for studying protein localization within the nucleus and nucleolus.
  • These reporters enable in vivo assessment of signal sequence function, independent of cellular context.

Purpose of the Study:

  • To present a standardized protocol for quantifying nuclear and nucleolar protein accumulation using NLS- and NoLS-fusion reporters.
  • To establish a robust framework for reliable measurement of NLS- and NoLS-mediated localization.

Main Methods:

  • The protocol integrates plasmid construction, cell transfection, and controlled protein expression.
  • It includes standardized imaging and computational analysis to minimize artifacts and improve reproducibility.
  • The workflow focuses on NLS- and NoLS-fusion reporters for tracking protein localization.

Main Results:

  • The developed protocol provides a standardized workflow for measuring nuclear and nucleolar accumulation.
  • It enhances the reproducibility and reliability of quantifying protein localization mediated by NLS and NoLS.
  • The framework allows for artifact reduction in localization studies.

Conclusions:

  • This standardized protocol offers a robust basis for quantifying NLS- and NoLS-mediated protein localization.
  • It facilitates the comparison of nuclear transport mechanisms across different studies.
  • The method supports reliable in vivo evaluation of signal sequence function.