This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Giachetti, C Articles by Semler, B L Search for Related Content PubMed PubMed Citation Articles by Giachetti, C Articles by Semler, B L

 Previous Article  |  Next Article 

J Virol. 1992 October; 66(10): 6045-6057

cis-acting lesions targeted to the hydrophobic domain of a poliovirus membrane protein involved in RNA replication.

Department of Microbiology and Molecular Genetics, College of Medicine, University of California, Irvine 92717.

ABSTRACT

The structural requirements of the hydrophobic domain contained in poliovirus polypeptide 3AB were studied by using a molecular genetic approach in combination with an in vitro biochemical analysis. We report here the generation and analysis of deletion, insertion, and amino acid replacement mutations aimed at decreasing the hydrophobic character of the domain. Our results indicated that the hydrophobicity of this region of 3AB is necessary to maintain normal viral RNA synthesis. However, in vitro membrane association assays of the mutated proteins did not establish a direct correlation between 3AB membrane association and viral RNA synthesis. Some of the lethal mutations we engineered produced polyproteins with abnormal P2- and P3-processing capabilities due to an alteration in the normal cleavage order of the polyprotein. A detailed analysis of these mutants suggests that P2 is not the major precursor for polypeptides 2A and 2BC and that P2 protein products are derived from P2-P3-containing precursors (most likely P2-P3 or P2-3AB). Such precursors are likely to result from primary polyprotein cleavage events that initiate a proteolytic cascade not previously documented. Our results also indicated that the function provided by the hydrophobic domain of 3AB cannot be provided in trans. We discuss the implications of these results on the formation of limited-diffusion replication complexes as a means of sequestering P2- and P3-region polypeptides required for RNA synthesis and protein processing.

This article has been cited by other articles:

• de Sessions, P. F., Dobrikova, E., Gromeier, M. (2007). Genetic Adaptation to Untranslated Region-Mediated Enterovirus Growth Deficits by Mutations in the Nonstructural Proteins 3AB and 3CD. J. Virol. 81: 8396-8405 [Abstract] [Full Text]  
• Yin, J., Liu, Y., Wimmer, E., Paul, A. V. (2007). Complete protein linkage map between the P2 and P3 non-structural proteins of poliovirus. J. Gen. Virol. 88: 2259-2267 [Abstract] [Full Text]  
• Strauss, D. M., Wuttke, D. S. (2007). Characterization of Protein-Protein Interactions Critical for Poliovirus Replication: Analysis of 3AB and VPg Binding to the RNA-Dependent RNA Polymerase. J. Virol. 81: 6369-6378 [Abstract] [Full Text]  
• Liu, Y., Franco, D., Paul, A. V., Wimmer, E. (2007). Tyrosine 3 of Poliovirus Terminal Peptide VPg(3B) Has an Essential Function in RNA Replication in the Context of Its Precursor Protein, 3AB. J. Virol. 81: 5669-5684 [Abstract] [Full Text]  
• Wessels, E., Duijsings, D., Lanke, K. H. W., van Dooren, S. H. J., Jackson, C. L., Melchers, W. J. G., van Kuppeveld, F. J. M. (2006). Effects of Picornavirus 3A Proteins on Protein Transport and GBF1-Dependent COP-I Recruitment. J. Virol. 80: 11852-11860 [Abstract] [Full Text]  
• Lee, H., Liu, Y., Mejia, E., Paul, A. V., Wimmer, E. (2006). The C-Terminal Hydrophobic Domain of Hepatitis C Virus RNA Polymerase NS5B Can Be Replaced with a Heterologous Domain of Poliovirus Protein 3A. J. Virol. 80: 11343-11354 [Abstract] [Full Text]  
• Wessels, E., Notebaart, R. A., Duijsings, D., Lanke, K., Vergeer, B., Melchers, W. J. G., van Kuppeveld, F. J. M. (2006). Structure-Function Analysis of the Coxsackievirus Protein 3A: IDENTIFICATION OF RESIDUES IMPORTANT FOR DIMERIZATION, VIRAL RNA REPLICATION, AND TRANSPORT INHIBITION. J. Biol. Chem. 281: 28232-28243 [Abstract] [Full Text]  
• Wessels, E., Duijsings, D., Notebaart, R. A., Melchers, W. J. G., van Kuppeveld, F. J. M. (2005). A Proline-Rich Region in the Coxsackievirus 3A Protein Is Required for the Protein To Inhibit Endoplasmic Reticulum-to-Golgi Transport. J. Virol. 79: 5163-5173 [Abstract] [Full Text]  
• Cornell, C. T., Brunner, J. E., Semler, B. L. (2004). Differential Rescue of Poliovirus RNA Replication Functions by Genetically Modified RNA Polymerase Precursors. J. Virol. 78: 13007-13018 [Abstract] [Full Text]  
• Choe, S. S., Kirkegaard, K. (2004). Intracellular Topology and Epitope Shielding of Poliovirus 3A Protein. J. Virol. 78: 5973-5982 [Abstract] [Full Text]  
• Grubman, M. J., Baxt, B. (2004). Foot-and-Mouth Disease. Clin. Microbiol. Rev. 17: 465-493 [Abstract] [Full Text]  
• Teterina, N. L., Rinaudo, M. S., Ehrenfeld, E. (2003). Strand-Specific RNA Synthesis Defects in a Poliovirus with a Mutation in Protein 3A. J. Virol. 77: 12679-12691 [Abstract] [Full Text]  
• Egger, D., Bienz, K. (2002). Recombination of Poliovirus RNA Proceeds in Mixed Replication Complexes Originating from Distinct Replication Start Sites. J. Virol. 76: 10960-10971 [Abstract] [Full Text]  
• van Kuppeveld, F. J. M., van den Hurk, P. J. J. C., Schrama, I. W. J., Galama, J. M. D., Melchers, W. J. G. (2002). Trans-complementation of a genetic defect in the coxsackie B3 virus 2B protein. J. Gen. Virol. 83: 341-350 [Abstract] [Full Text]  
• Dodd, D. A., Giddings, T. H. Jr., Kirkegaard, K. (2001). Poliovirus 3A Protein Limits Interleukin-6 (IL-6), IL-8, and Beta Interferon Secretion during Viral Infection. J. Virol. 75: 8158-8165 [Abstract] [Full Text]  
• Núñez, J. I., Baranowski, E., Molina, N., Ruiz-Jarabo, C. M., Sánchez, C., Domingo, E., Sobrino, F. (2001). A Single Amino Acid Substitution in Nonstructural Protein 3A Can Mediate Adaptation of Foot-and-Mouth Disease Virus to the Guinea Pig. J. Virol. 75: 3977-3983 [Abstract] [Full Text]  
• Genovese, D, Catone, S, Farah, M., Gambacorta, A, Fiore, L (1999). Isolation and biological characterization of 3(2H)-isoflavene-resistant and -dependent poliovirus type 2 Sabin mutants. J. Gen. Virol. 80: 157-167 [Abstract]  
• Towner, J. S., Mazanet, M. M., Semler, B. L. (1998). Rescue of Defective Poliovirus RNA Replication by 3AB-Containing Precursor Polyproteins. J. Virol. 72: 7191-7200 [Abstract] [Full Text]  
• Xiang, W., Cuconati, A., Hope, D., Kirkegaard, K., Wimmer, E. (1998). Complete Protein Linkage Map of Poliovirus P3 Proteins: Interaction of Polymerase 3Dpol with VPg and with Genetic Variants of 3AB. J. Virol. 72: 6732-6741 [Abstract] [Full Text]  
• Towner, J. S., V. Ho, T., Semler, B. L. (1996). Determinants of Membrane Association for Poliovirus Protein 3AB. J. Biol. Chem. 271: 26810-26818 [Abstract] [Full Text]  
• Lama, J., Sanz, M. A., Rodrguez, P. L. (1995). A Role for 3AB Protein in Poliovirus Genome Replication. J. Biol. Chem. 270: 14430-14438 [Abstract] [Full Text]  
• Novak, J E, Kirkegaard, K (1994). Coupling between genome translation and replication in an RNA virus.. Genes Dev. 8: 1726-1737 [Abstract]