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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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Updated: Jan 31, 2026

Subcellular Fractionation of Primary Chronic Lymphocytic Leukemia Cells to Monitor Nuclear/Cytoplasmic Protein Trafficking
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Nuclear/Cytoplasmic Fractionation to Study Hippo Effectors.

Maria Chatzifrangkeskou1, Eric O'Neill2

  • 1Department of Oncology, University of Oxford, Oxford, UK.

Methods in Molecular Biology (Clifton, N.J.)
|December 20, 2018
PubMed
Summary

Researchers developed a fast method to separate nuclear and cytoplasmic fractions, preserving protein integrity. This technique aids in studying Hippo protein shuttling in response to cellular cues.

Keywords:
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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Hippo proteins translocate between the nucleus and cytoplasm rapidly in response to various stimuli.
  • Understanding this translocation is crucial for cell growth and tissue homeostasis.
  • Existing methods for fraction separation can be time-consuming and may compromise protein integrity.

Purpose of the Study:

  • To develop and present a novel, rapid experimental procedure for separating nuclear and cytoplasmic protein fractions.
  • To ensure the preservation of protein integrity and protein-protein complexes during fractionation.

Main Methods:

  • A simple and fast protocol for the separation of nuclear and cytoplasmic fractions was established.
  • The procedure was designed to maintain the integrity of proteins and their complexes.

Main Results:

  • The described method allows for quick separation of cellular compartments.
  • Crucially, the procedure maintains the integrity of proteins and protein-protein complexes.
  • This indicates the method's suitability for studying dynamic protein translocation events.

Conclusions:

  • A robust and efficient method for nuclear-cytoplasmic fractionation is presented.
  • This technique facilitates the study of Hippo protein shuttling and other signaling pathways.
  • The method's ability to preserve molecular integrity broadens its applicability in cell biology research.