Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 103, Issue 44, Pages 16496-16501Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0605835103
Keywords
cell entry; hemolysis; membrane penetration; reoviridae
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Funding
- NIAID NIH HHS [F31 AI 064142, R01 AI 46440, F31 AI064142, R01 AI046440] Funding Source: Medline
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During cell entry, reovirus particles with a diameter of 70-80 nm must penetrate the cellular membrane to access the cytoplasm. The mechanism of penetration, without benefit of membrane fusion, is not well characterized for any such nonenveloped animal virus. Lysis of RBCs is an in vitro assay for the membrane perforation activity of reovirus; however, the mechanism of lysis has been unknown. In this report, osmotic-protection experiments using PEGs of different sizes revealed that reovirus-incluced lysis of RBCs occurs osmotically, after formation of small size-selective lesions or pores. Consistent results were obtained by monitoring leakage of fluorophore-tagged dextrans from the interior of resealed RBC ghosts. Gradient fractionations showed that whole virus particles, as Well as the myristoylated fragment mu 1N that is released from particles, are recruited to RBC membranes in association with pore formation. We propose that formation of small pores is a discrete, intermediate step in the reovirus membrane-penetration pathway, which may be shared by other nonenveloped animal viruses.
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