This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow E-mail this article to a friend
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 Barco, A.
Right arrow Articles by Carrasco, L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Barco, A.
Right arrow Articles by Carrasco, L.

 Previous Article  |  Next Article 

J Virol, May 1998, p. 3560-3570, Vol. 72, No. 5
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Identification of Regions of Poliovirus 2BC Protein That Are Involved in Cytotoxicity

Angel Barco* and Luis Carrasco

Centro de Biología Molecular (CSIC-UAM), Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain

Received 22 September 1997/Accepted 20 January 1998

The expression of poliovirus 2BC protein in yeast and mammalian cells leads to a number of metabolic and morphological alterations, such as growth inhibition, intracellular membrane proliferation, blockade of the exocytic pathway, and enhanced membrane permeability. Yeast cells that express poliovirus 2BC in an inducible manner were used to identify the regions of 2BC implicated in the modifications of these cellular functions. Several 2BC deletion mutants were generated to define the minimal portion of 2BC required to alter these activities. Additional deletion mutants that were obtained by random mutagenesis followed by selection in yeast cells provided new insights into the structure and mechanism of action of 2BC. The activity responsible for membrane proliferation is located in 2C, while the activities responsible for membrane permeabilization and inhibition of the exocytic pathway are located in 2B. Several regions of 2B and 2C required for the different functions of 2BC were identified. Thus, the integrity of the N termini of both 2B and 2C is necessary for 2BC-induced cytotoxicity. It is also possible to separate the different cellular alterations provoked by 2BC by the use of several 2BC variants. Deletion of amino acids 52 to 65 in 2B generates a 2BC deletion variant, 2bCDelta AvrII, that still blocks yeast growth but is unable to enhance membrane permeability or to inhibit the exocytic pathway. On the other hand, 2Bc128*.32b and 2Bc128*.3c, which contain only 73 and 77 amino acids of 2B, interfere with yeast division and enhance membrane permeability but affect the exocytic pathway only weakly and do not induce membrane proliferation. Our findings indicate that Saccharomyces cerevisiae represents a useful model system to analyze the functions of poliovirus 2BC and show the feasibility of separating the activities assigned to this protein.

* Corresponding author. Mailing address: Centro de Biología Molecular (CSIC-UAM), Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain. Phone: 34-1-397 8450. Fax: 34-1-397 4799. E-mail: abarco{at}trasto.cbm.uam.es.

J Virol, May 1998, p. 3560-3570, Vol. 72, No. 5
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Slobodskaya, O., Laarman, A., Spaan, W. J. M. (2008). Intracellular Restriction of a Productive Noncytopathic Coronavirus Infection. J. Virol. 82: 451-460 [Abstract] [Full Text]  
  • Salako, M. A., Carter, M. J., Kass, G. E. N. (2006). Coxsackievirus Protein 2BC Blocks Host Cell Apoptosis by Inhibiting Caspase-3. J. Biol. Chem. 281: 16296-16304 [Abstract] [Full Text]  
  • Teterina, N. L., Levenson, E., Rinaudo, M. S., Egger, D., Bienz, K., Gorbalenya, A. E., Ehrenfeld, E. (2006). Evidence for Functional Protein Interactions Required for Poliovirus RNA Replication. J. Virol. 80: 5327-5337 [Abstract] [Full Text]  
  • Belov, G. A., Fogg, M. H., Ehrenfeld, E. (2005). Poliovirus Proteins Induce Membrane Association of GTPase ADP-Ribosylation Factor. J. Virol. 79: 7207-7216 [Abstract] [Full Text]  
  • Harris, J. R., Racaniello, V. R. (2005). Amino Acid Changes in Proteins 2B and 3A Mediate Rhinovirus Type 39 Growth in Mouse Cells. J. Virol. 79: 5363-5373 [Abstract] [Full Text]  
  • Moffat, K., Howell, G., Knox, C., Belsham, G. J., Monaghan, P., Ryan, M. D., Wileman, T. (2005). Effects of Foot-and-Mouth Disease Virus Nonstructural Proteins on the Structure and Function of the Early Secretory Pathway: 2BC but Not 3A Blocks Endoplasmic Reticulum-to-Golgi Transport. J. Virol. 79: 4382-4395 [Abstract] [Full Text]  
  • de Jong, A. S., Melchers, W. J. G., Glaudemans, D. H. R. F., Willems, P. H. G. M., van Kuppeveld, F. J. M. (2004). Mutational Analysis of Different Regions in the Coxsackievirus 2B Protein: REQUIREMENTS FOR HOMO-MULTIMERIZATION, MEMBRANE PERMEABILIZATION, SUBCELLULAR LOCALIZATION, AND VIRUS REPLICATION. J. Biol. Chem. 279: 19924-19935 [Abstract] [Full Text]  
  • Harris, J. R., Racaniello, V. R. (2003). Changes in Rhinovirus Protein 2C Allow Efficient Replication in Mouse Cells. J. Virol. 77: 4773-4780 [Abstract] [Full Text]  
  • Jurgens, C., Flanegan, J. B. (2003). Initiation of Poliovirus Negative-Strand RNA Synthesis Requires Precursor Forms of P2 Proteins. J. Virol. 77: 1075-1083 [Abstract] [Full Text]  
  • Agirre, A., Barco, A., Carrasco, L., Nieva, J. L. (2002). Viroporin-mediated Membrane Permeabilization. PORE FORMATION BY NONSTRUCTURAL POLIOVIRUS 2B PROTEIN. J. Biol. Chem. 277: 40434-40441 [Abstract] [Full Text]  
  • Carette, J. E., van Lent, J., MacFarlane, S. A., Wellink, J., van Kammen, A. (2002). Cowpea Mosaic Virus 32- and 60-Kilodalton Replication Proteins Target and Change the Morphology of Endoplasmic Reticulum Membranes. J. Virol. 76: 6293-6301 [Abstract] [Full Text]  
  • de Jong, A. S., Schrama, I. W. J., Willems, P. H. G. M., Galama, J. M. D., Melchers, W. J. G., van Kuppeveld, F. J. M. (2002). Multimerization reactions of coxsackievirus proteins 2B, 2C and 2BC: a mammalian two-hybrid analysis. J. Gen. Virol. 83: 783-793 [Abstract] [Full Text]  
  • Neznanov, N., Kondratova, A., Chumakov, K. M., Angres, B., Zhumabayeva, B., Agol, V. I., Gudkov, A. V. (2001). Poliovirus Protein 3A Inhibits Tumor Necrosis Factor (TNF)-Induced Apoptosis by Eliminating the TNF Receptor from the Cell Surface. J. Virol. 75: 10409-10420 [Abstract] [Full Text]  
  • Sanz, M. A., Carrasco, L. (2001). Sindbis Virus Variant with a Deletion in the 6K Gene Shows Defects in Glycoprotein Processing and Trafficking: Lack of Complementation by a Wild-Type 6K Gene in trans. J. Virol. 75: 7778-7784 [Abstract] [Full Text]  
  • Deora, A., Ratner, L. (2001). Viral Protein U (Vpu)-Mediated Enhancement of Human Immunodeficiency Virus Type 1 Particle Release Depends on the Rate of Cellular Proliferation. J. Virol. 75: 6714-6718 [Abstract] [Full Text]  
  • Chang, Y.-S., Liao, C.-L., Tsao, C.-H., Chen, M.-C., Liu, C.-I, Chen, L.-K., Lin, Y.-L. (1999). Membrane Permeabilization by Small Hydrophobic Nonstructural Proteins of Japanese Encephalitis Virus. J. Virol. 73: 6257-6264 [Abstract] [Full Text]  


Copyright © 2010 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»