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Cholesterol removal by methyl-beta-cyclodextrin inhibits poliovirus entry
1Department of Microbiology and Immunology, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205, USA.
Journal of Virology
|December 13, 2003
Summary
Poliovirus entry into cells depends on cellular cholesterol levels, not detergent-insoluble microdomains. Cholesterol depletion inhibits genome delivery after viral capsid conformational changes, indicating a distinct entry pathway.
Area of Science:
- Virology
- Cell Biology
- Membrane Biology
Background:
- Poliovirus entry involves conformational changes of viral particles upon receptor binding.
- The precise cellular membranes and mechanisms for poliovirus genome delivery remain unclear.
- Detergent-insoluble microdomains (DIMs) are implicated in other picornavirus entry pathways.
Purpose of the Study:
- To investigate the role of detergent-insoluble microdomains (DIMs) and cellular cholesterol in poliovirus entry.
- To determine if poliovirus utilizes DIMs for infection.
- To elucidate the stage of entry affected by cholesterol depletion.
Main Methods:
- Disruption of DIMs using methyl-beta-cyclodextrin (MbetaCD) to deplete cellular cholesterol.
- Fractionation of cellular membranes to localize poliovirus receptor (PVR) and capsid proteins.
- Assessment of viral infection inhibition and RNA delivery after MbetaCD treatment.
Main Results:
- MbetaCD treatment inhibited poliovirus infection, suggesting cholesterol's importance.
- Poliovirus and PVR localized to detergent-soluble membrane fractions, not DIMs, during entry.
- MbetaCD treatment inhibited RNA delivery post-135S particle formation, independent of DIMs.
Conclusions:
- Cellular cholesterol levels are critical for poliovirus genome delivery.
- Poliovirus entry and genome delivery utilize a pathway distinct from those dependent on DIM integrity.
- The findings highlight a novel cholesterol-dependent mechanism in viral RNA release.