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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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Nuclear Export of mRNA02:31

Nuclear Export of mRNA

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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 Export01:42

Nuclear Export

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The nucleus restricts several proteins within and allows others to pass. The restricted proteins possess a nuclear retention sequence or NRS, anchoring them to the nuclear lamins and preventing their transport to the cytosol. The non-restricted proteins, after their synthesis, are transported to their site of action, such as the cytosol or other organelles, with the help of nuclear export signals or NES.
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Nuclear Localization Signals and Import01:46

Nuclear Localization Signals and Import

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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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Regulation of Nuclear Protein Sorting01:45

Regulation of Nuclear Protein Sorting

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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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Protein Complex Assembly02:41

Protein Complex Assembly

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Proteins can form homomeric complexes with another unit of the same protein or heteromeric complexes with different types.  Most protein complexes self-assemble spontaneously via ordered pathways, while some proteins need assembly factors that guide their proper assembly. Despite the crowded intracellular environment, proteins usually interact with their correct partners and form functional complexes.
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Related Experiment Video

Updated: Sep 27, 2025

In Vitro Nuclear Assembly Using Fractionated Xenopus Egg Extracts
04:49

In Vitro Nuclear Assembly Using Fractionated Xenopus Egg Extracts

Published on: September 2, 2008

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Nuclear Pore Complex Assembly Using Xenopus Egg Extract.

Guillaume Holzer1, Wolfram Antonin2

  • 1Institute of Biochemistry and Molecular Cell Biology, Medical School, RWTH Aachen University, Aachen, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|April 12, 2022
PubMed
Summary
This summary is machine-generated.

Xenopus egg extract enables in vitro study of nuclear pore complex assembly. Researchers can analyze protein function using antibody depletion and assays like Western blot or immunofluorescence.

Keywords:
Annulate lamellaeImmunofluorescenceInterphase egg extractNuclear assemblyNuclear pore complexWestern blotXenopus laevis

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A Cell-Free Assay Using Xenopus laevis Embryo Extracts to Study Mechanisms of Nuclear Size Regulation
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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Xenopus egg extract is a valuable in vitro system for studying complex cellular processes.
  • Nuclear pore complex (NPC) assembly is crucial for nucleocytoplasmic transport and cellular function.

Purpose of the Study:

  • To describe the methodology for obtaining and utilizing interphase Xenopus egg extract.
  • To detail methods for analyzing nuclear pore complex assembly in vitro.
  • To demonstrate protein function analysis via antibody-mediated depletion.

Main Methods:

  • Preparation of interphase Xenopus egg extract.
  • In vitro assays for nuclear pore complex assembly.
  • Western blot and immunofluorescence for analysis.
  • Antibody-mediated protein depletion for functional studies.

Main Results:

  • Established a reliable protocol for Xenopus egg extract preparation.
  • Successfully visualized and analyzed nuclear pore complex assembly in vitro.
  • Demonstrated the efficacy of antibody-mediated depletion for assessing protein function in NPC assembly.

Conclusions:

  • Interphase Xenopus egg extract is a robust model for studying nuclear pore complex assembly.
  • Western blot and immunofluorescence are effective analytical tools for this process.
  • Antibody-mediated depletion provides a powerful method to investigate protein roles in cellular mechanisms.