Rotavirus infection of cells in culture induces activation of RhoA and changes in the actin and tubulin cytoskeleton.

Rotavirus infection induces an increase in [Ca(2+)](cyto), which in turn may affect the distribution of the cytoskeleton proteins in the infected cell. Changes in microfilaments, including the formation of stress fibers, were observed starting at 0.5 h.p.i. using fluorescent phalloidin. Western blot...

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Main Authors: Jose Luis Zambrano, Orlando Sorondo, Ana Alcala, Esmeralda Vizzi, Yuleima Diaz, Marie Christine Ruiz, Fabian Michelangeli, Ferdinando Liprandi, Juan E Ludert
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2012-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC3474729?pdf=render
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author Jose Luis Zambrano
Orlando Sorondo
Ana Alcala
Esmeralda Vizzi
Yuleima Diaz
Marie Christine Ruiz
Fabian Michelangeli
Ferdinando Liprandi
Juan E Ludert
author_facet Jose Luis Zambrano
Orlando Sorondo
Ana Alcala
Esmeralda Vizzi
Yuleima Diaz
Marie Christine Ruiz
Fabian Michelangeli
Ferdinando Liprandi
Juan E Ludert
author_sort Jose Luis Zambrano
collection DOAJ
description Rotavirus infection induces an increase in [Ca(2+)](cyto), which in turn may affect the distribution of the cytoskeleton proteins in the infected cell. Changes in microfilaments, including the formation of stress fibers, were observed starting at 0.5 h.p.i. using fluorescent phalloidin. Western blot analysis indicated that RhoA is activated between 0.5 and 1 h.p.i. Neither the phosphorylation of RhoA nor the formation of stress fibers were observed in cells infected with virions pre-treated with an anti-VP5* non-neutralizing mAb, suggesting that RhoA activation is stimulated by the interaction of the virus with integrins forming the cell receptor complex. In addition, the structure of the tubulin cytoskeleton was also studied. Alterations of the microtubules were evident starting at 3 h.p.i. and by 7 h.p.i. when microtubules were markedly displaced toward the periphery of the cell cytoplasm. Loading of rotavirus-infected cells with either a Ca(2+) chelator (BAPTA) or transfection with siRNAs to silence NSP4, reversed the changes observed in both the microfilaments and microtubules distribution, but not the appearance of stress fibers. These results indicate that alterations in the distribution of actin microfilaments are initiated early during infection by the activation of RhoA, and that latter changes in the Ca(2+) homeostasis promoted by NSP4 during infection may be responsible for other alterations in the actin and tubulin cytoskeleton.
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spelling doaj.art-7b4d0b6f7b7b45d4bee0fee081aaf44f2022-12-21T17:25:38ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-01710e4761210.1371/journal.pone.0047612Rotavirus infection of cells in culture induces activation of RhoA and changes in the actin and tubulin cytoskeleton.Jose Luis ZambranoOrlando SorondoAna AlcalaEsmeralda VizziYuleima DiazMarie Christine RuizFabian MichelangeliFerdinando LiprandiJuan E LudertRotavirus infection induces an increase in [Ca(2+)](cyto), which in turn may affect the distribution of the cytoskeleton proteins in the infected cell. Changes in microfilaments, including the formation of stress fibers, were observed starting at 0.5 h.p.i. using fluorescent phalloidin. Western blot analysis indicated that RhoA is activated between 0.5 and 1 h.p.i. Neither the phosphorylation of RhoA nor the formation of stress fibers were observed in cells infected with virions pre-treated with an anti-VP5* non-neutralizing mAb, suggesting that RhoA activation is stimulated by the interaction of the virus with integrins forming the cell receptor complex. In addition, the structure of the tubulin cytoskeleton was also studied. Alterations of the microtubules were evident starting at 3 h.p.i. and by 7 h.p.i. when microtubules were markedly displaced toward the periphery of the cell cytoplasm. Loading of rotavirus-infected cells with either a Ca(2+) chelator (BAPTA) or transfection with siRNAs to silence NSP4, reversed the changes observed in both the microfilaments and microtubules distribution, but not the appearance of stress fibers. These results indicate that alterations in the distribution of actin microfilaments are initiated early during infection by the activation of RhoA, and that latter changes in the Ca(2+) homeostasis promoted by NSP4 during infection may be responsible for other alterations in the actin and tubulin cytoskeleton.http://europepmc.org/articles/PMC3474729?pdf=render
spellingShingle Jose Luis Zambrano
Orlando Sorondo
Ana Alcala
Esmeralda Vizzi
Yuleima Diaz
Marie Christine Ruiz
Fabian Michelangeli
Ferdinando Liprandi
Juan E Ludert
Rotavirus infection of cells in culture induces activation of RhoA and changes in the actin and tubulin cytoskeleton.
PLoS ONE
title Rotavirus infection of cells in culture induces activation of RhoA and changes in the actin and tubulin cytoskeleton.
title_full Rotavirus infection of cells in culture induces activation of RhoA and changes in the actin and tubulin cytoskeleton.
title_fullStr Rotavirus infection of cells in culture induces activation of RhoA and changes in the actin and tubulin cytoskeleton.
title_full_unstemmed Rotavirus infection of cells in culture induces activation of RhoA and changes in the actin and tubulin cytoskeleton.
title_short Rotavirus infection of cells in culture induces activation of RhoA and changes in the actin and tubulin cytoskeleton.
title_sort rotavirus infection of cells in culture induces activation of rhoa and changes in the actin and tubulin cytoskeleton
url http://europepmc.org/articles/PMC3474729?pdf=render
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