Document Detail

Infectious rotavirus enters cells by direct cell membrane penetration, not by endocytosis.
MedLine Citation:
PMID:  2831376     Owner:  NLM     Status:  MEDLINE    
Rotaviruses are icosahedral viruses with a segmented, double-stranded RNA genome. They are the major cause of severe infantile infectious diarrhea. Rotavirus growth in tissue culture is markedly enhanced by pretreatment of virus with trypsin. Trypsin activation is associated with cleavage of the viral hemagglutinin (viral protein 3 [VP3]; 88 kilodaltons) into two fragments (60 and 28 kilodaltons). The mechanism by which proteolytic cleavage leads to enhanced growth is unknown. Cleavage of VP3 does not alter viral binding to cell monolayers. In previous electron microscopic studies of infected cell cultures, it has been demonstrated that rotavirus particles enter cells by both endocytosis and direct cell membrane penetration. To determine whether trypsin treatment affected rotavirus internalization, we studied the kinetics of entry of infectious rhesus rotavirus (RRV) into MA104 cells. Trypsin-activated RRV was internalized with a half-time of 3 to 5 min, while nonactivated virus disappeared from the cell surface with a half-time of 30 to 50 min. In contrast to trypsin-activated RRV, loss of nonactivated RRV from the cell surface did not result in the appearance of infection, as measured by plaque formation. Endocytosis inhibitors (sodium azide, dinitrophenol) and lysosomotropic agents (ammonium chloride, chloroquine) had a limited effect on the entry of infectious virus into cells. Purified trypsin-activated RRV added to cell monolayers at pH 7.4 medicated 51Cr, [14C]choline, and [3H]inositol released from prelabeled MA104 cells. This release could be specifically blocked by neutralizing antibodies to VP3. These results suggest that MA104 cell infection follows the rapid entry of trypsin-activated RRV by direct cell membrane penetration. Cell membrane penetration of infectious RRV is initiated by trypsin cleavage of VP3. Neutralizing antibodies can inhibit this direct membrane penetration.
K T Kaljot; R D Shaw; D H Rubin; H B Greenberg
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of virology     Volume:  62     ISSN:  0022-538X     ISO Abbreviation:  J. Virol.     Publication Date:  1988 Apr 
Date Detail:
Created Date:  1988-04-15     Completed Date:  1988-04-15     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  0113724     Medline TA:  J Virol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1136-44     Citation Subset:  IM    
Department of Medicine, Stanford University School of Medicine, California 95305.
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MeSH Terms
Ammonium Chloride / pharmacology
Azides / pharmacology
Cell Line
Cell Membrane / microbiology*
Cell Membrane Permeability
Chloroquine / pharmacology
Dinitrophenols / pharmacology
Hemagglutinins, Viral / metabolism
Rotavirus / drug effects,  growth & development,  physiology*
Sodium Azide
Trypsin / pharmacology*
Grant Support
Reg. No./Substance:
0/Azides; 0/Dinitrophenols; 0/Hemagglutinins, Viral; 12125-02-9/Ammonium Chloride; 26628-22-8/Sodium Azide; 54-05-7/Chloroquine; EC

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