This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Tian, P.
Right arrow Articles by Schilling, W. P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Tian, P.
Right arrow Articles by Schilling, W. P.

 Previous Article  |  Next Article 

J. Virol., Sep 1995, 5763-5772, Vol 69, No. 9
Copyright © 1995, American Society for Microbiology

The rotavirus nonstructural glycoprotein NSP4 mobilizes Ca2+ from the endoplasmic reticulum

P Tian, MK Estes, Y Hu, JM Ball, CQ Zeng and WP Schilling
Division of Molecular Virology, Baylor College of Medicine, Houston, Texas 77030, USA.

We previously reported that expression of rotavirus nonstructural glycoprotein NSP4 is responsible for an increase in cytosolic free Ca2+ concentration ([Ca2+]i) in Spodoptera frugiperda (Sf9) insect cells (P. Tian, Y. Hu, W. P. Schilling, D. A. Lindsay, J. Eiden, and M. K. Estes, J. Virol. 68:251-257, 1994). The purpose of the present study was to determine the mechanism by which NSP4 causes an increase in [Ca2+]i by measuring the permeability of the cytoplasmic and endoplasmic reticulum (ER) membranes in recombinant-baculovirus-infected Sf9 cells. No obvious change in plasmalemma permeability to divalent cations was observed in cells expressing NSP4 compared with that in cells expressing another rotaviral glycoprotein (VP7) when the influx of Ba2+, a Ca2+ surrogate, was monitored. The basal Ca2+ permeability of the internal Ca2+ store was evaluated by measuring the release of Ca2+ induced by ionomycin, a Ca2+ ionophore, or thapsigargin, an inhibitor of the ER Ca(2+)-ATPase pump, following suspension of the cells in Ca(2+)-free extracellular buffer. Releasable Ca2+ decreased with time to a greater extent in cells expressing NSP4 compared with that in cells expressing VP7, suggesting that NSP4 increases the basal Ca2+ permeability of the ER membrane. To determine the possible mechanism by which NSP4 increases ER permeability, purified NSP4 protein or a 22- amino-acid synthetic peptide consisting of residues 114 to 135 (NSP4(114-135) was added exogenously to noninfected Sf9 cells during measurement of [Ca2+]i. Both NSP4 and the NSP4(114-135 peptide produced a time-dependent increase in [Ca2+]i that was attenuated by prior inhibition of phospholipase C with U-73122. Pretreatment of the cells with thapsigargin completely blocked the increase in [Ca2+]i produced by NSP4(114-135, but the peptide only partially reduced the change in [Ca2+]i produced by thapsigargin. No changes in [Ca2+]i were seen in cells treated with control peptides. These results suggest that (i) exogenous NSP4 increases [Ca2+]i through the activation of phospholipase C, (ii) Ca2+ release by exogenous NSP4 is from a store that is a subset of the thapsigargin-sensitive compartment, and (iii) amino acid residues 114 to 135 of NSP4 are sufficient for this activity. In contrast to exogenous NSP4, the mechanism by which endogenously expressed NSP4 increases [Ca2+]1 appears to be unrelated to phospholipase C, since no effect of U-73122 was seen on the elevated [Ca2+]1 in cells expressing NSP4 and exogenously applied NSP4(114-135) caused a further increase in [Ca2+]1 in cells expressing NSP4 protein.(ABSTRACT TRUNCATED AT 400 WORDS)


This article has been cited by other articles:

  • Diaz, Y., Chemello, M. E., Pena, F., Aristimuno, O. C., Zambrano, J. L., Rojas, H., Bartoli, F., Salazar, L., Chwetzoff, S., Sapin, C., Trugnan, G., Michelangeli, F., Ruiz, M. C. (2008). Expression of Nonstructural Rotavirus Protein NSP4 Mimics Ca2+ Homeostasis Changes Induced by Rotavirus Infection in Cultured Cells. J. Virol. 82: 11331-11343 [Abstract] [Full Text]  
  • Lin, Y.-P., Kao, C.-L., Chang, S.-Y., Taniguchi, K., Hung, P.-Y., Lin, H.-C., Huang, L.-M., Huang, H.-H., Yang, J.-Y., Lee, C.-N. (2008). Determination of Human Rotavirus VP6 Genogroups I and II by Reverse Transcription-PCR. J. Clin. Microbiol. 46: 3330-3337 [Abstract] [Full Text]  
  • Sen, A., Sen, N., Mackow, E. R. (2007). The Formation of Viroplasm-Like Structures by the Rotavirus NSP5 Protein Is Calcium Regulated and Directed by a C-Terminal Helical Domain. J. Virol. 81: 11758-11767 [Abstract] [Full Text]  
  • Martin-Latil, S., Mousson, L., Autret, A., Colbere-Garapin, F., Blondel, B. (2007). Bax Is Activated during Rotavirus-Induced Apoptosis through the Mitochondrial Pathway. J. Virol. 81: 4457-4464 [Abstract] [Full Text]  
  • Mascarenhas, J. D. P., Leite, J. P. G., Lima, J. C., Heinemann, M. B., Oliveira, D. S., Araujo, I. T., Soares, L. S., Gusmao, R. H. P., Gabbay, Y. B., Linhares, A. C. (2007). Detection of a neonatal human rotavirus strain with VP4 and NSP4 genes of porcine origin. J Med Microbiol 56: 524-532 [Abstract] [Full Text]  
  • Lorin-Nebel, C., Xing, J., Yan, X., Strange, K. (2007). CRAC channel activity in C. elegans is mediated by Orai1 and STIM1 homologues and is essential for ovulation and fertility. J. Physiol. 580: 67-85 [Abstract] [Full Text]  
  • Yan, X., Xing, J., Lorin-Nebel, C., Estevez, A. Y., Nehrke, K., Lamitina, T., Strange, K. (2006). Function of a STIM1 Homologue in C. elegans: Evidence that Store-operated Ca2+ Entry Is Not Essential for Oscillatory Ca2+ Signaling and ER Ca2+ Homeostasis. JGP 128: 443-459 [Abstract] [Full Text]  
  • Parr, R. D., Storey, S. M., Mitchell, D. M., McIntosh, A. L., Zhou, M., Mir, K. D., Ball, J. M. (2006). The Rotavirus Enterotoxin NSP4 Directly Interacts with the Caveolar Structural Protein Caveolin-1. J. Virol. 80: 2842-2854 [Abstract] [Full Text]  
  • Lopez, T., Camacho, M., Zayas, M., Najera, R., Sanchez, R., Arias, C. F., Lopez, S. (2005). Silencing the Morphogenesis of Rotavirus. J. Virol. 79: 184-192 [Abstract] [Full Text]  
  • Ramig, R. F. (2004). Pathogenesis of Intestinal and Systemic Rotavirus Infection. J. Virol. 78: 10213-10220 [Full Text]  
  • Boshuizen, J. A., Rossen, J. W. A., Sitaram, C. K., Kimenai, F. F. P., Simons-Oosterhuis, Y., Laffeber, C., Buller, H. A., Einerhand, A. W. C. (2004). Rotavirus Enterotoxin NSP4 Binds to the Extracellular Matrix Proteins Laminin-{beta}3 and Fibronectin. J. Virol. 78: 10045-10053 [Abstract] [Full Text]  
  • Iturriza Gomara, M., Kang, G., Mammen, A., Jana, A. K., Abraham, M., Desselberger, U., Brown, D., Gray, J. (2004). Characterization of G10P[11] Rotaviruses Causing Acute Gastroenteritis in Neonates and Infants in Vellore, India. J. Clin. Microbiol. 42: 2541-2547 [Abstract] [Full Text]  
  • Griffin, S. D. C., Harvey, R., Clarke, D. S., Barclay, W. S., Harris, M., Rowlands, D. J. (2004). A conserved basic loop in hepatitis C virus p7 protein is required for amantadine-sensitive ion channel activity in mammalian cells but is dispensable for localization to mitochondria. J. Gen. Virol. 85: 451-461 [Abstract] [Full Text]  
  • Bergqvist, A., Sundstrom, S., Dimberg, L. Y., Gylfe, E., Masucci, M. G. (2003). The Hepatitis C Virus Core Protein Modulates T Cell Responses by Inducing Spontaneous and Altering T-cell Receptor-triggered Ca2+ Oscillations. J. Biol. Chem. 278: 18877-18883 [Abstract] [Full Text]  
  • Mirazimi, A., Magnusson, K.-E., Svensson, L. (2003). A cytoplasmic region of the NSP4 enterotoxin of rotavirus is involved in retention in the endoplasmic reticulum. J. Gen. Virol. 84: 875-883 [Abstract] [Full Text]  
  • Ding, W., Albrecht, B., Kelley, R. E., Muthusamy, N., Kim, S.-J., Altschuld, R. A., Lairmore, M. D. (2002). Human T-Cell Lymphotropic Virus Type 1 p12I Expression Increases Cytoplasmic Calcium To Enhance the Activation of Nuclear Factor of Activated T Cells. J. Virol. 76: 10374-10382 [Abstract] [Full Text]  
  • Enouf, V., Langella, P., Commissaire, J., Cohen, J., Corthier, G. (2001). Bovine Rotavirus Nonstructural Protein 4 Produced by Lactococcus lactis Is Antigenic and Immunogenic. Appl. Environ. Microbiol. 67: 1423-1428 [Abstract] [Full Text]  
  • Tafazoli, F., Zeng, C. Q., Estes, M. K., Magnusson, K.-E., Svensson, L. (2001). NSP4 Enterotoxin of Rotavirus Induces Paracellular Leakage in Polarized Epithelial Cells. J. Virol. 75: 1540-1546 [Abstract] [Full Text]  
  • Zhang, M., Zeng, C. Q.-Y., Morris, A. P., Estes, M. K. (2000). A Functional NSP4 Enterotoxin Peptide Secreted from Rotavirus-Infected Cells. J. Virol. 74: 11663-11670 [Abstract] [Full Text]  
  • Lee, C.-N., Wang, Y.-L., Kao, C.-L., Zao, C.-L., Lee, C.-Y., Chen, H.-N. (2000). NSP4 Gene Analysis of Rotaviruses Recovered from Infected Children with and without Diarrhea. J. Clin. Microbiol. 38: 4471-4477 [Abstract] [Full Text]  
  • Brunet, J.-P., Jourdan, N., Cotte-Laffitte, J., Linxe, C., Géniteau-Legendre, M., Servin, A., Quéro, A.-M. (2000). Rotavirus Infection Induces Cytoskeleton Disorganization in Human Intestinal Epithelial Cells: Implication of an Increase in Intracellular Calcium Concentration. J. Virol. 74: 10801-10806 [Abstract] [Full Text]  
  • Browne, E. P., Bellamy, A. R., Taylor, J. A. (2000). Membrane-destabilizing activity of rotavirus NSP4 is mediated by a membrane-proximal amphipathic domain. J. Gen. Virol. 81: 1955-1959 [Abstract] [Full Text]  
  • Obert, G., Peiffer, I., Servin, A. L. (2000). Rotavirus-Induced Structural and Functional Alterations in Tight Junctions of Polarized Intestinal Caco-2 Cell Monolayers. J. Virol. 74: 4645-4651 [Abstract] [Full Text]  
  • Brunet, J.-P., Cotte-Laffitte, J., Linxe, C., Quero, A.-M., Géniteau-Legendre, M., Servin, A. (2000). Rotavirus Infection Induces an Increase in Intracellular Calcium Concentration in Human Intestinal Epithelial Cells: Role in Microvillar Actin Alteration. J. Virol. 74: 2323-2332 [Abstract] [Full Text]  
  • Pérez, J. F., Ruiz, M.-C., Chemello, M. E., Michelangeli, F. (1999). Characterization of a Membrane Calcium Pathway Induced by Rotavirus Infection in Cultured Cells. J. Virol. 73: 2481-2490 [Abstract] [Full Text]  
  • Xu, A., Bellamy, A. R., Taylor, J. A. (1998). BiP (GRP78) and Endoplasmin (GRP94) Are Induced following Rotavirus Infection and Bind Transiently to an Endoplasmic Reticulum-Localized Virion Component. J. Virol. 72: 9865-9872 [Abstract] [Full Text]  
  • Zhang, M., Zeng, C. Q.-Y., Dong, Y., Ball, J. M., Saif, L. J., Morris, A. P., Estes, M. K. (1998). Mutations in Rotavirus Nonstructural Glycoprotein NSP4 Are Associated with Altered Virus Virulence. J. Virol. 72: 3666-3672 [Abstract] [Full Text]  
  • Coulson, B. S., Londrigan, S. L., Lee, D. J. (1997). Rotavirus contains integrin ligand sequences and a disintegrin-like domain that are implicated in virus entry into cells. Proc. Natl. Acad. Sci. USA 94: 5389-5394 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»