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[Mg2+-dependent interaction of DNA with eukaryotic cells]
Abstract:
Interaction of DNA with eukaryotic cells under conditions similar to those providing DNA adsorption onto liposomes was studied. It was revealed that mouse fibroblasts (line A9) and myeloma cells bind phage and plasmid DNA in 0.3 M sucrose solution containing Mg2+-ions. Additional pretreatment of the cells by trypsin did not affect DNA adsorption efficiency. The major part of the adsorbed DNA recovered by salt treatment of the cells, but 10-15% of DNA was found to be irreversible. Up to 50% of the irreversibly bound DNA molecules retain their linear size after treatment of cells with DNAse I. Efficiencies of DNA adsorption and irreversibly binding depend on the concentration of Mg2+ in the medium. The process of DNA irreversible binding is not inhibited by drugs affecting cell metabolism. It is assumed that DNA adsorbs onto the phospholipid domains of the cell membrane, and part of the adsorbed DNA is taken up into the interior of the cells.
Insights
Eukaryotic cells, including mouse fibroblasts and myeloma cells, can bind DNA in a specific solution. A portion of this bound DNA is irreversibly attached, suggesting cellular uptake mechanisms.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Context:
- Investigates DNA interaction with eukaryotic cells under conditions mimicking liposome adsorption.
- Utilizes mouse fibroblasts (line A9) and myeloma cells for experimental models.
- Employs a 0.3 M sucrose solution with magnesium ions (Mg2+) to facilitate DNA binding.
Purpose:
- To study the mechanisms of DNA adsorption and irreversible binding to eukaryotic cells.
- To determine the influence of magnesium ion concentration on DNA-cell interactions.
- To explore the fate and stability of adsorbed DNA within the cellular environment.
Summary:
- Mouse fibroblasts and myeloma cells bind phage and plasmid DNA in a Mg2+-containing sucrose solution.
- Adsorption efficiency is not affected by trypsin pretreatment but depends on Mg2+ concentration.
- 10-15% of adsorbed DNA binds irreversibly, with a significant portion retaining its size after DNase I treatment, suggesting potential cellular internalization.
Impact:
- Provides insights into DNA-cell membrane interactions and potential DNA internalization pathways.
- Highlights the role of Mg2+ in modulating DNA binding to eukaryotic cells.
- Suggests that a portion of extracellular DNA may become stably associated with or enter cells, independent of metabolic processes.