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Influence of polycations on the interaction between poliovirus multistranded ribonucleic acid and HeLa cells
Abstract:
In contrast to single-stranded viral ribonucleic acid (RNA), poliovirus multistranded RNA (RI-RNA) is poorly adsorbed to suspended HeLa cells in the absence of polycations. In the presence of 20 mug of diethylaminoethyl (DEAE) dextran per ml and other polycations, 90% or more of the RI-RNA is adsorbed to HeLa cells within 2 min at 37 C. However, only 30% of the RI-RNA label penetrates into the cells independent of the concentration of DEAE dextran applied. DEAE dextran is adsorbed almost quantitatively to HeLa cells within 3 min at 37 C. Most of the DEAE dextran remains bound to the cell membrane and available for binding of RI-RNA for 15 min at 37 C.
Insights
Poliovirus multistranded RNA (RI-RNA) poorly adsorbs to cells alone. Diethylaminoethyl (DEAE) dextran enhances RI-RNA adsorption to HeLa cells, but cell penetration remains limited.
Area of Science:
- Virology
- Cell Biology
- Biochemistry
Background:
- Single-stranded viral RNA differs in cellular adsorption compared to poliovirus multistranded RNA (RI-RNA).
- Polycations are known to influence the interaction between nucleic acids and cells.
Purpose of the Study:
- To investigate the effect of polycations, specifically diethylaminoethyl (DEAE) dextran, on the adsorption of poliovirus RI-RNA to HeLa cells.
- To determine the extent of RI-RNA penetration into HeLa cells in the presence of DEAE dextran.
Main Methods:
- Experiments involved incubating suspended HeLa cells with poliovirus RI-RNA in the presence and absence of DEAE dextran at 37°C.
- Adsorption and cell penetration of RI-RNA were quantified using radiolabeling.
- The binding of DEAE dextran to HeLa cells was also assessed over time.
Main Results:
- In the absence of polycations, RI-RNA showed poor adsorption to HeLa cells.
- The addition of DEAE dextran (20 µg/ml) resulted in over 90% adsorption of RI-RNA to HeLa cells within 2 minutes.
- Despite high adsorption, only about 30% of the RI-RNA label penetrated the cells, irrespective of DEAE dextran concentration.
- DEAE dextran itself was rapidly and almost quantitatively adsorbed to HeLa cells, remaining available for RI-RNA binding for at least 15 minutes.
Conclusions:
- Diethylaminoethyl (DEAE) dextran significantly enhances the adsorption of poliovirus RI-RNA to HeLa cells.
- While DEAE dextran facilitates RI-RNA binding to the cell surface, it does not proportionally increase the intracellular penetration of the viral RNA.
- The cell membrane binding of DEAE dextran is stable, suggesting its potential as a tool for studying RNA-cell interactions.