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Two modes of entry of reovirus particles into L cells
Abstract:
Evidence is presented supporting the hypothesis that reovirus intermediate subviral particles (ISVP), which show increased infectivity relative to intact virions, can gain entry into host L cells by two alternative pathways. One pathway is by the process of viropexis, involving phagocytic vacuoles. A second entry pathway is via direct penetration of the plasma membrane of the cell, without involvement of a phagocytic vacuole. Using electron microscopy, a kinetic analysis of the uptake process was carried out. Results indicate that at 37 degrees C ISVP gain entry into host cells primarily by direct entry, although viropexis also occurs, while intact virions gain entry by viropexis almost exclusively. A second line of experimental evidence consistent with the idea that ISVP can 'melt' their way through the plasma membrane is provided by studies on the release of pre-loaded radioactive 51Cr from host cells following infection. 51Cr release data demonstrate that infection with ISVP leads to an immediate increased leakiness of the cell plasma membrane, whereas no such increase takes place following infection with an equivalent number of intact virions. This demonstrates that ISVP can interact with the plasma membrane of the cell in a manner which is qualitatively different from the interaction between intact virions and the plasma membrane. The ability of ISVP to directly penetrate the plasma membrane of the host cell, which intact virions apparently cannot do, could explain the decreased duration of the eclipse phase, as well as the increased infectivity of ISVP, relative to that observed for infection with intact virions.
Insights
Reovirus intermediate subviral particles (ISVP) infect cells through direct plasma membrane penetration, unlike intact virions that use viropexis. This ISVP entry mechanism explains their higher infectivity and shorter eclipse phase.
Area of Science:
- Virology
- Cell Biology
- Molecular Biology
Background:
- Reovirus intermediate subviral particles (ISVP) exhibit higher infectivity than intact virions.
- Understanding viral entry mechanisms is crucial for controlling viral infections.
Purpose of the Study:
- To investigate the entry pathways of reovirus ISVP into host L cells.
- To compare the entry mechanisms of ISVP with those of intact reovirus virions.
Main Methods:
- Electron microscopy was used for kinetic analysis of viral uptake.
- Radioactive 51Cr release assay was employed to assess plasma membrane integrity.
Main Results:
- ISVP primarily enter cells via direct plasma membrane penetration at 37°C, with some viropexis observed.
- Intact reovirus virions predominantly enter cells through viropexis.
- ISVP infection causes immediate plasma membrane leakiness, unlike intact virions.
Conclusions:
- ISVP possess a unique ability to directly penetrate host cell plasma membranes.
- This direct entry mechanism of ISVP contributes to their increased infectivity and reduced eclipse phase compared to intact virions.