Abstract
Rotavirus (RV) is a triple-protein-layered icosahedral virus, for which studies have established that the two outer-layer proteins, viral protein 4 (VP4) and viral protein 7 (VP7), are required for viral infectivity (1,2). VP7, a glycoprotein, is the major component of the outer-layer, but its role in viral entry is unclear. VP4 forms dimers extending out from the VP7-coated viral surface (3,4) and have been shown to be a determinant of host range and virulence, and is directly involved in cell attachment and RV entry into cells (5–8). Proteolytic cleavage of VP4 into two noncovalently associated subunits, VP8* and VP5* (2,9,10), significantly enhances viral infectivity (11–13).
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Bridger, J. C. and Woode, G. N. (1976) Characterization of two particle types of calf rotavirus. J. Gen. Virol. 31, 245–250.
Estes, M. K., Graham, D. Y., Smith, E. M., and Gerba, C. P. (1979) Rotavirus stability and inactivation. J. Gen. Virol. 43, 403–409.
Prasad, B. V. V., Burns, J. W., Marietta, E. Estes, M. K., and Chiu, W. (1990) Localization of VP4 neutralization sites in rotavirus by three-dimensional cryo-electron microscopy. Nature 343, 476–479.
Shaw, A. L., Rothnagel, R., Chen, D., Ramig, R. F., Chiu, W., and Prasad, B. V. V. (1993) Three-dimensional visualization of the rotavirus hemagglutinin structure. Cell 74, 693–701.
Greenberg, H. B., Flores, J., Kalica, A. R., Wyatt, R. G., and Jones, R. (1983) Gene coding assignments for growth restriction, neutralization and subgroup specificities of the W and DS-1 strains of human rotavirus. J. Gen. Virol. 64, 313–320.
Kaljot, K. T., Shaw, R. D., Rubin, D. H., and Greenberg, H. B. (1988) Infectious rotavirus enters cells by direct cell membrane penetration, not by endocytosis. J. Virol. 62, 1136–1144.
Offit, P. A., Blavat, G., Greenberg, H. B., and Clark, H. F. (1986) Molecular basis of rotavirus virulence: role of gene segment 4. J. Virol. 57, 46–49.
Ruggeri, F. M. and Greenberg, H. B. (1991) Antibodies to the trypsin cleavage peptide VP8 neutralize rotavirus by inhibiting binding of virions to target cells in culture. J. Virol. 65, 2211–2219.
Espejo, R. T., Lopez, S., and Arias, C. (1981) Structural polypeptides of simian rotavirus SA11 and the effect of trypsin. J. Virol. 37, 156–160.
Lopez, S., Arias, C. F., Bell, J. R., Strauss, J. H., and Espejo, R. T. (1985) Primary structure of the cleavage site associated with trypsin enhancement of rotavirus SA11 infectivity. Virology 144, 11–19.
Babiuk, L. A., Mohammed, K., Spence, L., Fauvel, M., and Petro, R. (1977) Rotavirus isolation and cultivation in the presence of trypsin. J. Clin. Microbiol. 6, 610–617.
Barnett, B. B., Spendlove, R. S., and Clark, M. L. (1979) Effect of enzymes on rotavirus infectivity. J. Clin. Microbiol. 10, 111–113.
Clark, S. M., Roth, J. R., Clark, M. L., Barnett, B. B., and Spendlove, R. S. (1981) Trypsin enhancement of rotavirus infectivity: mechanism of enhancement. J. Virol. 39, 816–822.
Fukuhara, N., Yoshie, O., Kitaoka, S., and Konno, T. (1988) Role of VP3 in human rotavirus internalization after target cell attachment via VP7. J. Virol. 62, 2209–2218.
Nandi, P., Charpilienne, A., and Cohen, J. (1992) Interaction of rotavirus particles with liposomes. J. Virol. 66, 3363–3367.
Ruiz, M. C., Alonso, T. S., Charpilienne, A., Vasseur, M., Michelangeli, F., Cohen, J., and Alvarado, F. (1994) Rotavirus interaction with isolated membrane vesicles. J. Virol. 68, 4009–4016.
Falconer, M. M., Gilbert, J. M., Roper, A. M., Greenberg, H. B., and Gavora, J. S. (1995) Rotavirus-induced fusion-from-without in tissue culture cells. J. Virol. 69, 5582–5591.
Carrasco, L. (1994) Entry of animal viruses and macromolecules into cells. FEBS Lett. 350, 151–154.
Cuadras, M. A., Arias, C. F., and Lopez, S. (1997) Rotaviruses induce an early membrane permabilization of MA104 cells and do not require a low intracellular Ca2+ concentration to initiate their replication cycle. J. Virol. 71, 9065–9074.
Liprandi, F., Moros, Z., Gerder, M., Ludert, J. E., Pujol, F. H., Ruiz, M. C., Michelangeli, F., Charpilienne, A., and Cohen, J. (1997) Productive penetration of rotavirus in cultured cells induces coentry of the translation inhibitor α-sarcin. Virology 237, 430–438.
Brigotti, M., Rambelli, F., Zamboni, M., Montanaro, L., and Sperti, L. (1989) Effect of α-sarcin and ribosome-inactivating proteins on the interaction of elongation factos with ribosomes. Biochem. J. 257, 723–727.
Endo, Y. and Wool, I. G. (1982) The site of action of α-sarcin on eukaryotic ribosome. J. Biol. Chem. 257, 9054–9060.
Gilbert, J. M. and Greenberg, H. B. (1997) Virus-like particle induced fusion-from-without in tissue culture cells; role of outer-layer proteins VP4 and VP7. J. Virol. 71, 4555–4563.
Crawford, S. E., Labbé, M., Cohen, J., Burroughs, M. H., Zhou, Y. J., and Estes, M. K. (1994) Characterization of virus-like particles produced by the expression of rotavirus capsid proteins in insect cells. J. Virol. 68, 5915–5922.
Gilbert, J. M. and Greenberg, H. B. (1998) Cleavage of rhesus rotavirus VP4 after arginine 247 is essential for rotavirus-like particle-induced fusion-from-without. J. Virol. 72, 5323–5327.
Mackow, E. R., Shaw, R. D., Matsui, S. M., Vo, P. T., Dang, M.-N., and Greenberg, H. B. (1988) The rhesus rotavirus gene encoding protein VP3: Location of amino acids involved in homologous and heterologous rotavirus neutralization and identification of a putative fusion region. Proc. Natl. Acad. Sci. USA 85, 645–649.
Buckland, R. and Wild, F. (1989) Leucine zipper motif extends. Nature 338, 547–548.
Mendez, E., Arias, C. F., and Lopez, S. (1996) Interactions between the two surface proteins of rotavirus may alter the receptor-binding specificity of the virus. J. Virol. 70, 1218–1222.
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© 2000 Humana Press Inc., Totowa, NJ
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Gilbert, J.M., Greenberg, H.B. (2000). Rotavirus Entry into Tissue Culture Cells. In: Gray, J., Desselberger, U. (eds) Rotaviruses. Methods in Molecular Medicine™, vol 34. Humana Press. https://doi.org/10.1385/1-59259-078-0:67
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DOI: https://doi.org/10.1385/1-59259-078-0:67
Publisher Name: Humana Press
Print ISBN: 978-0-89603-736-6
Online ISBN: 978-1-59259-078-0
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