Publicado
La proteína viral VO6 de rotavirus coinmunoprecipita con HSC70 en células MA104
The viral protein VP6 of rotavirus coinmunoprecipated with HSC70 in cels MA104
Palabras clave:
infección por rotavirus, vaccinia, reoviridae, rhesus (macaca mulatta). (es)rotavirus infections, vaccinia, reoviridae, rhesus (macaca mulatta) (en)
Introducción: los rotavirus son la principal causa de gastroenteritis en niños menores de cinco años. Hasta el momento se desconoce cómo entran estos virus a la célula huésped. La proteína de choque térmico, HSC70 se ha visto involucrada en la entrada del virus, sin embargo no se han relacionado bien todos los sitios de unión entre las proteínas virales del rotavirus y la HSC70.
Objetivos: el propósito de este trabajo fue determinar cuáles proteínas virales del rotavirus son responsables de la unión de la proteína de choque térmico, HSC70.
Material y métodos: se realizaron ensayos de coinmunoprecipitación utilizando lisados de células infectadas así como proteínas virales recombinantes. Estos ensayos se revelaron mediante SDS-PAGE y Western blot utilizando anticuerpos policlonales que reconocen las proteínas del rotavirus.
Resultados. En los experimentos realizados se encontró que la proteína viral VP6 coinmunoprecipita con HSC70 en las células MA104 durante la infección con rotavirus. La VP6, producida con vaccinia recombinante en MA104, también coinmunoprecipitó con HSC70.
Conclusión. La proteína VP6 de rotavirus interacciona con la proteína HSC70 en lisados de células MA104 infectadas con rotavirus. La unión de la VP6 recombinante a la HSC70 sugiere que la interacción de estas proteínas es independiente de otras proteínas virales.
Introduction: Rotaviruses are the main cause of gastroenteritis in children under five years old. At this moment is unknown how these viruses entry to host cell. The heat shock cognate protein HSC70 has been involved in rotavirus cell entry. However, the relationship between rotavirus proteins and HSC70 has not been established.
Objectives: the purpose of this work was to determine wich rotaviruses proteins interact with HSC70.
Material and methods: coimmunoprecipitation assays were done on a rotavirus infected supernatant and recombinant viral proteins. These assays were revealed by SDS-PAGE and Western blot using policlonal antibodys against rotavirus structural proteins.
Results: data show that viral protein VP6 coimmunoprecipated with HSC70 in MA104 cells during infection. Recombinant vaccinia of VP6 produced in MA104 cells also coimmunoprecipitated with HSC70.
Conclusions: these results suggest the interaction between VP6 and HSC70 during rotavirus infection. Furthermore, this union may be independent of other rotaviruses proteins.
Referencias
Ball JM, Tian P , Zeng CQY, Morris AP, Estes MK, Age-dependent diarrhea induced by a rotaviral nonstructural glycoprotein. Science. 1996;272:101-104.
Dubois E, Guyader Le F, Haugarreau L, Kopecka H, Cormier M, Pommepuy M. "Molecular Epidemiological Survey of Rotaviruses in Sewage by Reverse Transcriptase Seminested PCR and Restriction Fragment Length Polymorphism Assay". Appl. Environ Microbiol. 1997;63: 1794-1800
Estes MK. "Rotaviruses and their Replication". Fields, B.N. Knipe, D.M. and Howley, eds. En Virology, Philadelphia: Lippencott Raven, 1996; 1625-1655.
Granoff A, Webster R. "Rotaviruses" Encyclopedia of Virology, San Diego, USA, Academic Press, 2da. Edición.1999;1576-1578.
Leite J, Alfieri AA, Woods PA, Glass RI, Gentsch JR. "Rotavirus G and P types circulating in brazil: characterization by RT-PCR, probe hybridization, and sequence analysis". Arch Virol. 1996;141: 2365-2374.
Lundgren O, Peregrin AT, Persson K, Kordasti S, Uhnoo I, Svensson L. Role of the enteric nervous system in the fluid and electrolyte secretion of rotavirus diarrhea. Science. 2000;287:491-495.
Guerrero CA, Bouyssounade D, Zarate S, Isa P, López T, Espinosa R, Romero P, Méndez E, López S, Arias CF. "Heat Shock Cognate Protein 70 Is Involved in Rotavirus Cell Entry". J. Virol. 2002; 76: 4096-4102.
Zárate S, Cuadras MA, Espinosa R, Romero P, Juárez KO, Camacho-Nuez M, Arias CF, López S. Interaction of Rotaviruses with HSC70 during Cell Entry Is Mediated by VP5. J Virol. 2003; 77 (13): 7254-7260
Cripe T, Delos S, Estes P, Garcea R. "In Vivo and In Vitro Association of Hsc70 with Polyomavirus Capsid Proteins". J. Virol. 1995;69:7807- 7813.
Laemmli UK. " Cleavage of structural proteins during the assembly of the head of bacteriophage t4". Nature 1970;227:680-685.
Chappell TG, Welch WJ, Schlossman DM, Palter KB, Schlesinger MJ, Rothman JE. Uncoating ATPaes is a member of the 70 kilodalton family of stress proteins. Cell. 1986;45:3-13
Chou Ch, Forouhar F, Yeh Y, Hui-Lin Shr, Wang Ch, Chwan-Deng Hsiao. Crystal Structure of the C-terminal 10-kDa Subdomain of HSC70. J. Biol. Chem. 2003; 278:30311-30316
Flaherty KM, Mackay DB, Kabsch W, Holmes KC. Similarity of the three-dimensional structures of actin and the ATPase fragment of a 70-kDa heat shock cognate protein. Proc. Natl. Acad. USA. 1991;88:5041-5045
Frydman J, Harti FU. Molecular chaperone functions of hsp70 and hsp60 in protein folding. The Biology of heat shock proteins and molecular chaperones. Cold spring Harbor Laboratory Press:1994;251-283.
Arias CF, Isa P, Guerrero CA, Mendez E, Zarate S, López T, López S. Molecular biology of rotavirus cell entry. Arch. Med. Res. 2002 ;33: 356- 36
Guerrero CA, Zarate S, Corkidi G, López S, Arias C. Biochemical characterization of rotavirus receptor in MA104 cells. J Virol. 2000; 74;93-102.
Méndez E, López S, Cuadras MA, Romero P, Arias CF. Entry of rotaviruses is a multistep process. Virology. 1999;263:450-459.
Sánchez-San Martin C, Lopez T, Arias CF, Lopez S. Characterization of rotavirus cell entry. J. Virol. 2004;78, 2310- 2318.
Zárate S, Espinosa R, Romero P, Méndez E, Arias CF, López S. The VP5 domain of VP4 can mediate the attachment of rotaviruses to cells. J. Virol. 2000. 74:593-599
Isa P, Realpe M, Romero P, López S, Arias CF. Rotavirus RRV associates with lipid membrane microdomains during cell entry. Virology. 2004;322:370-381
Burns J, Siadat-Pajouh, Krishnaney A, Greenberg H. Protective effect of rotavirus VP6-specific IgA monoclonal Antibodies that lack neutralizing activity. Science. 1996;272: 104-107
Jacobson RM. The current status of the rotavirus vaccine. Vaccine.1999; 17:1690-1699.
Choi A, Basu M, Mcneal M, Clements J, Ward R. Antibody-independent protection against rotavirus infection of mice stimulated by intranasal immunization with chimeric VP4 or VP6 protein. J. Virol. 1999;73:7574-7581.
Feng N, Lawton J, Gilgert J, Kuklin N, Vo P, Prasad B, Greenberg H. Inhibition of rotavirus replication by a non-neutralizing, rotavirus VP6-specific IgA mAb J Clin Invest.2002;109:1203-1213
Visitas a la página del resumen del artículo
Descargas
Cómo citar
Licencia
Derechos de autor 2005 Revista de la Facultad de Medicina
Esta obra está bajo una licencia internacional Creative Commons Atribución 4.0.
-
