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Related Experiment Videos

Aggregation of nucleosomes by divalent cations.

M de Frutos1, E Raspaud, A Leforestier

  • 1Laboratoire de Physique des Solides, Université de Paris Sud, 91405 Orsay Cedex, France. defrutos@lps.u-psud.fr

Biophysical Journal
|July 21, 2001
PubMed
Summary

Nucleosome core particles precipitate with divalent cations, forming phase diagrams similar to DNA. Charge reversal occurs with higher cation valency, impacting aggregate properties but not phase separation.

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Area of Science:

  • Biochemistry
  • Biophysics
  • Molecular Biology

Background:

  • Nucleosome core particles (NCPs) are fundamental units of DNA packaging.
  • Understanding NCP interactions is crucial for comprehending chromatin structure and function.
  • Cation-induced precipitation is a known phenomenon for nucleic acids and polyelectrolytes.

Purpose of the Study:

  • To investigate the precipitation conditions of NCPs induced by divalent cations.
  • To explore the relationship between cation concentration, NCP concentration, and phase separation.
  • To analyze the charge properties of NCP aggregates formed under different ionic conditions.

Main Methods:

  • Systematic variation of nucleosome core particle and cation concentrations.
  • Phase diagram construction to delineate precipitation and redissolution boundaries.

Related Experiment Videos

  • Electrophoretic mobility measurements to determine the effective charge of NCP aggregates.

    • Main Results:

    • NCP precipitation occurs above a specific divalent cation concentration threshold, followed by redissolution at higher concentrations.
    • The observed phase diagram resembles those of DNA and synthetic polyelectrolytes, suggesting cation condensation drives NCP interactions.
    • Aggregates formed with Mg(2+) remain negatively charged, while those with spermidine or spermine become positively charged, with charge reversal increasing with cation valency.
    • The effective charge sign did not influence the overall phase diagram.

    Conclusions:

    • NCP-NCP interactions are significantly influenced by cation condensation, similar to other polyanions.
    • The valency of counterions plays a critical role in determining the surface charge of NCP aggregates.
    • Theoretical models are needed to fully explain the observed charge reversal phenomenon in NCP aggregation.