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First published online 19 October 2004
doi: 10.1242/jcs.01425

Journal of Cell Science 117, 5509-5519 (2004)
Published by The Company of Biologists 2004
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Research Article

The rotavirus surface protein VP8 modulates the gate and fence function of tight junctions in epithelial cells

Porfirio Nava1, Susana López2, Carlos F. Arias2, Socorro Islas1 and Lorenza González-Mariscal1,*

1 Department of Physiology, Biophysics and Neuroscience, Center for Research and Advanced Studies (CINVESTAV), Mexico D.F. 07000, Mexico
2 Department of Developmental Genetics and Molecular Physiology, Institute of Biotechnology, National University of Mexico (UNAM), Cuernavaca, Morelos 62250, Mexico

* Author for correspondence (e-mail: lorenza{at}fisio.cinvestav.mx)

Accepted 30 July 2004

Rotaviruses constitute a major cause of diarrhea in young mammals. Rotaviruses utilize different integrins as cell receptors, therefore upon their arrival to the intestinal lumen their integrin receptors will be hidden below the tight junction (TJ), on the basolateral membrane. Here we have studied whether the rotavirus outer capsid proteins are capable of opening the paracellular space sealed by the TJ. From the outermost layer of proteins of the rotavirus, 60 spikes formed of protein VP4 are projected. VP4 is essential for virus-cell interactions and is cleaved by trypsin into peptides VP5 and VP8. Here we found that when these peptides are added to confluent epithelial monolayers (Madin-Darby canine kidney cells), VP8 is capable of diminishing in a dose dependent and reversible manner the transepithelial electrical resistance. VP5 exerted no effect. VP8 can also inhibit the development of newly formed TJs in a Ca-switch assay. Treatment with VP8 augments the paracellular passage of non-ionic tracers, allows the diffusion of a fluorescent lipid probe and the apical surface protein GP135, from the luminal to the lateral membrane, and triggers the movement of the basolateral proteins Na+-K+-ATPase, {alpha}{nu}ß3 integrin and ß1 integrin subunit, to the apical surface. VP8 generates a freeze-fracture pattern of TJs characterized by the appearance of loose end filaments, that correlates with an altered distribution of several TJ proteins. VP8 given orally to diabetic rats allows the enteral administration of insulin, thus indicating that it can be employed to modulate epithelial permeability.

Key words: Rotavirus, VP8, Tight junction, Occludin, Claudin, Drug delivery




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