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Protein Dynamics in Living Cells01:19

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Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
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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 Fast and Reliable Nuclei Extraction Method for Phytochrome Functional Analysis.

Ilse Villmann1, Akira Nagatani2, Mathias Zeidler3

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Methods in Molecular Biology (Clifton, N.J.)
|October 1, 2025
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Summary
This summary is machine-generated.

Phytochromes, plant light sensors, move between cytoplasm and nucleus to regulate growth. A new protocol isolates these compartments to identify their interaction partners and functions in each location.

Keywords:
NuclearNuclear importPhytochromeProtein extractioncytoplasmic partitioning

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Area of Science:

  • Plant biology
  • Molecular signaling

Background:

  • Phytochromes are crucial plant photoreceptors sensing red/far-red light.
  • Light-dependent nuclear translocation of phytochromes regulates plant development.
  • Emerging evidence suggests cytoplasmic roles for phytochromes.

Purpose of the Study:

  • To develop a protocol for isolating intact nuclei and cytoplasmic fractions from Arabidopsis thaliana seedlings.
  • To enable identification of phytochrome interaction partners in distinct cellular compartments.
  • To elucidate the differential roles of phytochromes in the cytoplasm versus the nucleus.

Main Methods:

  • Shearing of Arabidopsis thaliana seedlings.
  • Purification of intact nuclei using a Percoll gradient.
  • Separation of cytoplasmic and nuclear fractions for downstream analysis.

Main Results:

  • A protocol was established for rapid and straightforward isolation of nuclei and cytoplasm.
  • This method is applicable to both etiolated and light-exposed seedlings.
  • The isolated fractions facilitate identification of protein interaction partners.

Conclusions:

  • The developed protocol allows for compartment-specific analysis of phytochrome signaling.
  • It provides a foundation for understanding the distinct functions of phytochromes in the cytoplasm and nucleus.
  • This method aids in discovering novel phytochrome interaction partners and signaling pathways.