Abstract
In the present study, a homologous rotavirus, ECwt, infecting small intestinal villi isolated from ICR and BALB/c mice were used as a model for identifying cell-surface molecules involved in rotavirus entry. Small-intestinal villi were treated with anti-Hsc70, anti-PDI, anti-integrin β3 or anti-ERp57 antibodies or their corresponding F(ab’)2 fragments before inoculation with rotavirus ECwt, RRV or Wa. Pretreatment of villi decreased virus infectivity by about 50–100 % depending of the rotavirus strain, antibody structure and detection assay used. Similar results were obtained by treating viral inocula with purified proteins Hsc70, PDI or integrin β3 before inoculation of untreated villi. Rotavirus infection of villi proved to be sensitive to membrane-impermeant thiol/disulfide inhibitors such as DTNB and bacitracin, suggesting the involvement of a redox reaction in infection. The present results suggest that PDI, Hsc70 and integrin β3 are used by both homologous and heterologous rotaviruses during infection of isolated mouse villi.
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References
Abou-Jaoude G, Sureau C (2007) Entry of hepatitis delta virus requires the conserved cysteine residues of the hepatitis B virus envelope protein antigenic loop and is blocked by inhibitors of thiol-disulfide exchange. J Virol 81:13057–13066
Aoki ST, Settembre EC, Trask SD, Greenberg HB, Harrison SC, Dormitzer PR (2009) Structure of rotavirus outer-layer protein VP7 bound with a neutralizing Fab. Science 324:1444–1447
Arias CF, Lizano M, López S (1987) Synthesis in Escherichia coli and immunological characterization of a polypeptide containing the cleavage sites associated with trypsin enhancement of rotavirus SA11 infectivity. J Gen Virol 68:633–642
Mendez E, Arias CF, Lopez S (1993) Binding to sialic acids is not an essential step for the entry of animal rotaviruses to epithelial cells in culture. J Virol 67:5253–5259
Baker M, Prasad BV (2010) Rotavirus cell entry. Curr Top Microbiol Immunol 343:121–148
Beaulieu JF (1992) Differential expression of the VLA family of integrins along the crypt-villus axis in the human small intestine. J Cell Sci 102:427–436
Calderon MN, Guerrero CA, Acosta O, Lopez S, Arias CF (2012) Inhibiting rotavirus infection by membrane-impermeant thiol/disulfide exchange blockers and antibodies against protein disulfide isomerase. Intervirology 55:451–464
Ciarlet M, Ludert JE, Iturriza-Gómara M, Liprandi F, Gray JJ, Desselberger U, Estes MK (2002) Initial interaction of rotavirus strains with N-acetylneuraminic (sialic) acid residues on the cell surface correlates with VP4 genotype, not species of origin. J Virol 76:4087–4095
Crawford SE, Mukherjee SK, Estes MK, Lawton JA, Shaw AL, Ramig RF, Prasad BV (2001) Trypsin cleavage stabilizes the rotavirus VP4 spike. J Virol 75:6052–6061
Croyle MA, Walter E, Janich S, Roessler BJ, Amidon GL (1998) Role of integrin expression in adenovirus-mediated gene delivery to the intestinal epithelium. Hum Gene Ther 9:561–573
Danti P (2011) Enter the kill zone: initiation of death signaling during virus entry. Virology 411:316–324
Delmas O, Breton M, Sapin C, Bivic AL, Colard O, Trugnan G (2007) Heterogeneity of raft-type membrane microdomains associated with VP4, the rotavirus spike protein, in Caco-2 and MA 104 cells. J Virol 81:1610–1618
Graham KL, Fleming FE, Halasz P, Hewish MJ, Nagesha HS, Holmes IH, Takada Y, Coulson BS (2005) Rotaviruses interact with α4β7 and α4β1 integrins by binding the same integrin domains as natural ligands. J Gen Virol 86:3397–3408
Graham KL, Halasz P, Tan Y, Hewish MJ, Takada Y, Mackow ER, Robinson MK, Coulson BS (2003) Integrin-using rotaviruses bind α2β1 integrin α2 I domain via VP4 DGE sequence and recognize αxβ2 and ανβ3 by using VP7 during cell entry. J Virol 77:9969–9978
Gualtero DF, Guzmán F, Acosta O, Guerrero CA (2007) Amino acid domains 280–297 of VP6 and 531–554 of VP4 are implicated in heat shock cognate protein hsc70-mediated rotavirus infection. Arch Virol 152:2183–2196
Guerrero CA, Mendez E, Zárate S, Isa P, López S, Arias CF (2000) Integrin αvβ3 mediates rotavirus cell entry. Proc Natl Acad Sci USA 97:14644–14649
Guerrero CA, Santana AY, Acosta O (2010) Mouse intestinal villi as a model system for studies of rotavirus infection. J Virol Methods 168:22–30
Guglielmi KM, McDonald SM, Patton JT (2010) Mechanism of intraparticle synthesis of the rotavirus double-stranded RNA genome. J Biol Chem 285:18123–18128
Gutiérrez M, Isa P, Sánchez-San Martin C, Pérez-Vargas J, Espinosa R, Arias CF, López S (2010) Different rotavirus strains enter MA104 cells through different endocytic pathways: the role of clathrin-mediated endocytosis. J Virol 84:9161–9169
Halasz P, Holloway G, Coulson BS (2010) Death mechanisms in epithelial cells following rotavirus infection, exposure to inactivated rotavirus or genome transfection. J Gen Virol 91:2007–2018
Hamilton TE, McClane SJ, Baldwin S, Burke C, Patel H, Rombeau JL, Raper SE (1997) Efficient adenoviral-mediated murine neonatal small intestinal gene transfer is dependent on αv integrin expression. J Pediatr Surg 32:1695–1703
Harlow E, Lane D (1988) Antibodies: a laboratory manual. Cold Spring Harbor Laboratory, New York
Haselhorst T, Fleming FE, Dyason JC, Hartnell RD, Yu X, Holloway G, Santegoets K, Kiefel MJ, Blanchard H et al (2009) Sialic acid dependence in rotavirus host cell invasion. Nat Chem Biol 5:91–93
Helenius A (2007) Virus entry and uncoating. In: Knipe DM, Howley PM (eds) Fields virology, 5th edn. Lippincott, Philadephia
Isa P, Realpe M, Romero P, Lopez S, Arias CF (2004) Rotavirus RRV associates with lipid membrane microdomains during cell entry. Virology 322:370–381
Isa P, Gutierrez M, Arias CF, Lopez S (2008) Rotavirus cell entry. Future Med 3:135–146
Jain S, McGinnes LW, Morrison TG (2007) Thiol/disulfide exchange is required for membrane fusion directed by the Newcastle disease virus fusion protein. J Virol 81:2328–2339
Jordan PA, Gibbins JM (2006) Extracellular disulfide exchange and the regulation of cellular function. Antioxid Redox Signal 8:312–324
Keirstead ND, Coombs KM (1998) Absence of superinfection exclusion during asynchronous reovirus infections of mouse, monkey, and human cell lines. Virus Res 54:225–235
Kim ISTS, Babyonyshev M, Dormitzer PR, Harrison SC (2010) Effect of mutations in VP5* hydrophobic loops on rotavirus cell entry. J Virol 84:6200–6207
Lahav J, Wijnen EM, Hess O, Hamaia SW, Griffiths D, Makris M, Caballero CG, Essex DW, Farndale RW (2003) Enzymatically catalyzed disulfide exchange is required for platelet adhesion to collagen via integrin α2β1. Blood 102:2085–2092
Lopez S, Arias CF (2006) Early steps in rotavirus cell entry. Curr Top Microbiol Immunol 309:39–66
Lopez S, Arias CF (2004) Multistep entry of rotavirus into cells: a versaillesque dance. Trends Microbiol 12:271–278
Mainou BA, Dermody TS (2011) Src kinase mediates productive endocytic sorting of reovirus during cell entry. J Virol 85:3203–3213
Martín-Villa JM, Ferre-Lopez S, Lopez-Suárez JC, Corell A, Pérez-Blas M, Arnaiz-Villena A (1997) Cell surface phenotype and ultramicroscopic analysis of purified human enterocytes: a possible antigen-presenting cell in the intestine. Tissue Antigens 50:586–592
Maruri-Avidal L, López S, Arias CF (2008) Endoplasmic reticulum chaperones are involved in the morphogenesis of rotavirus infectious particles. J Virol 82:5368–5380
Mendez E, Lopez S, Cuadras MA, Romero P, Arias CF (1999) Entry of rotaviruses is a multistep process. Virology 263:450–459
Mirazimi A, Svensson L (1998) Carbohydrates facilitate correct disulfide bond formation and folding of rotavirus VP7. J Virol 72:3887–3892
O’Neil lS, Robinson A, Deering A, Ryan M, Fitzgerald DJ, Moran N (2000) The platelet integrin αIIbβ3 has an endogenous thiol isomerase activity. J Biol Chem
Parashar UD, Gibson CJ, Bresee JS, Glass RI (2006) Rotavirus and severe childhood diarrhea. Emerg Infect Dis 12:304–306
Pesavento JB, Crawford SE, Estes MK, Prasad BV (2006) Rotavirus proteins: structure and assembly. Curr Top Microbiol Immunol 309:189–219
Ryser HJ, Levy EM, Mandel R, DiSciullo GJ (1994) Inhibition of human immunodeficiency virus infection by agents that interfere with thiol-disulfide interchange upon virus-receptor interaction. Proc Natl Acad Sci USA 91:4559–4563
Simovich M, Hainsworth LN, Fields P, Umbreit JN, Conrad ME (2003) Localization of the iron transport proteins mobilferrin and DMT-1 in the duodenum: the surprising role of mucin. Am J Hematol 74:32–45
Stewart L, Ireton GC, Champoux JJ (1997) Reconstitution of human topoisomerase I by fragment complementation. J Mol Biol 269:355–357
Swiatkowska M, Szymanski J, Padula G, Cierniewski CS (2008) Interaction and functional association of protein disulfide isomerase with αVβ3 integrin on endothelial cells. FEBS J 275:1813–1823
Takada Y, Ye X, Simon S (2007) Protein family review. The integrins. Genome Biol 8:215
Yoder JD, Trask SD, Vo PT, Binka M, Feng N, Harrison SC, Greenberg HB, Dormitzer PR (2009) VP5* rearranges when rotavirus uncoats. J Virol 83:11372–11377
Zarate S, Cuadras MA, Espinosa R, Romero P, Juarez KO, Camacho-Nuez M, Arias CF, Lopez S (2003) Interaction of rotaviruses with Hsc70 during cell entry is mediated by VP5. J Virol 77:7254–7260
Zárate S, Espinosa R, Romero P, Arias CF, López S (2004) VP7 mediates the interaction of rotaviruses with integrin αvβ3 through a novel integrin-binding site. J Virol 78:10839–10847
Acknowledgments
This study was supported in part by a grant (No. 1101-405-20300, 193-2007) awarded by Colciencias to O. Acosta and C. A. Guerrero.
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Santana, A.Y., Guerrero, C.A. & Acosta, O. Implication of Hsc70, PDI and integrin αvβ3 involvement during entry of the murine rotavirus ECwt into small-intestinal villi of suckling mice. Arch Virol 158, 1323–1336 (2013). https://doi.org/10.1007/s00705-013-1626-6
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DOI: https://doi.org/10.1007/s00705-013-1626-6