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
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 Ahola, T.
Right arrow Articles by Auvinen, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ahola, T.
Right arrow Articles by Auvinen, P.

 Previous Article  |  Next Article 

Journal of Virology, August 2000, p. 6725-6733, Vol. 74, No. 15
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Effects of Palmitoylation of Replicase Protein nsP1 on Alphavirus Infection

Tero Ahola,1,dagger Pekka Kujala,1 Minna Tuittila,2 Titta Blom,1 Pirjo Laakkonen,1,Dagger Ari Hinkkanen,2 and Petri Auvinen1,*

Research Program in Cellular Biotechnology, Institute of Biotechnology, FIN-00014 University of Helsinki,1 and Department of Biochemistry and Pharmacy, Abo Akademi University, FIN-20521 Turku,2 Finland

Received 21 January 2000/Accepted 26 April 2000

The membrane-associated alphavirus RNA replication complex contains four virus-encoded subunits, the nonstructural proteins nsP1 to nsP4. Semliki Forest virus (SFV) nsP1 is hydrophobically modified by palmitoylation of cysteines 418 to 420. Here we show that Sindbis virus nsP1 is also palmitoylated on the same site (cysteine 420). When mutations preventing nsP1 palmitoylation were introduced into the genomes of these two alphaviruses, the mutant viruses remained viable and replicated to high titers, although their growth was slightly delayed. The subcellular distribution of palmitoylation-defective nsP1 was altered in the mutant: it no longer localized to filopodial extensions, and a fraction of it was soluble. The ultrastructure of the alphavirus replication sites appeared normal, and the localization of the other nonstructural proteins was unaltered in the mutants. In both wild-type- and mutant-virus-infected cells, SFV nsP3 and nsP4 could be extracted from membranes only by alkaline solutions whereas the nsP2-membrane association was looser. Thus, the membrane binding properties of the alphavirus RNA replication complex were not determined by the palmitoylation of nsP1. The nsP1 palmitoylation-defective alphaviruses produced normal plaques in several cell types, but failed to give rise to plaques in HeLa cells, although they induced normal apoptosis of these cells. The SFV mutant was apathogenic in mice: it caused blood viremia, but no infectious virus was detected in the brain.

* Corresponding author. Mailing address: Institute of Biotechnology, P.O. Box 56 (Viikinkaari 9), FIN-00014 University of Helsinki, Finland. Phone: 358-9-19158902. Fax: 358-9-19158952. E-mail: Petri.Auvinen{at}Helsinki.Fi.

dagger Present address: Institute for Molecular Virology, University of Wisconsin, Madison, WI 53706.

Dagger Present address: The Burnham Institute, La Jolla, CA 92037.

Journal of Virology, August 2000, p. 6725-6733, Vol. 74, No. 15
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Kiiver, K., Tagen, I., Zusinaite, E., Tamberg, N., Fazakerley, J. K., Merits, A. (2008). Properties of non-structural protein 1 of Semliki Forest virus and its interference with virus replication. J. Gen. Virol. 89: 1457-1466 [Abstract] [Full Text]  
  • Zusinaite, E., Tints, K., Kiiver, K., Spuul, P., Karo-Astover, L., Merits, A., Sarand, I. (2007). Mutations at the palmitoylation site of non-structural protein nsP1 of Semliki Forest virus attenuate virus replication and cause accumulation of compensatory mutations. J. Gen. Virol. 88: 1977-1985 [Abstract] [Full Text]  
  • Atasheva, S., Gorchakov, R., English, R., Frolov, I., Frolova, E. (2007). Development of Sindbis Viruses Encoding nsP2/GFP Chimeric Proteins and Their Application for Studying nsP2 Functioning. J. Virol. 81: 5046-5057 [Abstract] [Full Text]  
  • Spuul, P., Salonen, A., Merits, A., Jokitalo, E., Kaariainen, L., Ahola, T. (2007). Role of the Amphipathic Peptide of Semliki Forest Virus Replicase Protein nsP1 in Membrane Association and Virus Replication. J. Virol. 81: 872-883 [Abstract] [Full Text]  
  • Galbraith, S. E., Sheahan, B. J., Atkins, G. J. (2006). Deletions in the hypervariable domain of the nsP3 gene attenuate Semliki Forest virus virulence.. J. Gen. Virol. 87: 937-947 [Abstract] [Full Text]  
  • Frolova, E., Gorchakov, R., Garmashova, N., Atasheva, S., Vergara, L. A., Frolov, I. (2006). Formation of nsP3-Specific Protein Complexes during Sindbis Virus Replication.. J. Virol. 80: 4122-4134 [Abstract] [Full Text]  
  • Sawicki, D. L., Perri, S., Polo, J. M., Sawicki, S. G. (2006). Role for nsP2 Proteins in the Cessation of Alphavirus Minus-Strand Synthesis by Host Cells. J. Virol. 80: 360-371 [Abstract] [Full Text]  
  • Diatta, A., Piver, E., Collin, C., Vaudin, P., Pages, J.-C. (2005). Semliki Forest virus-derived virus-like particles: characterization of their production and transduction pathways. J. Gen. Virol. 86: 3129-3136 [Abstract] [Full Text]  
  • GREENE, I. P., PAESSLER, S., ANISHCHENKO, M., SMITH, D. R., BRAULT, A. C., FROLOV, I., WEAVER, S. C. (2005). VENEZUELAN EQUINE ENCEPHALITIS VIRUS IN THE GUINEA PIG MODEL: EVIDENCE FOR EPIZOOTIC VIRULENCE DETERMINANTS OUTSIDE THE E2 ENVELOPE GLYCOPROTEIN GENE. Am J Trop Med Hyg 72: 330-338 [Abstract] [Full Text]  
  • De, I., Fata-Hartley, C., Sawicki, S. G., Sawicki, D. L. (2003). Functional Analysis of nsP3 Phosphoprotein Mutants of Sindbis Virus. J. Virol. 77: 13106-13116 [Abstract] [Full Text]  
  • Tuittila, M., Hinkkanen, A. E. (2003). Amino acid mutations in the replicase protein nsP3 of Semliki Forest virus cumulatively affect neurovirulence. J. Gen. Virol. 84: 1525-1533 [Abstract] [Full Text]  
  • Salonen, A., Vasiljeva, L., Merits, A., Magden, J., Jokitalo, E., Kaariainen, L. (2003). Properly Folded Nonstructural Polyprotein Directs the Semliki Forest Virus Replication Complex to the Endosomal Compartment. J. Virol. 77: 1691-1702 [Abstract] [Full Text]  
  • Fazakerley, J. K., Boyd, A., Mikkola, M. L., Kaariainen, L. (2002). A Single Amino Acid Change in the Nuclear Localization Sequence of the nsP2 Protein Affects the Neurovirulence of Semliki Forest Virus. J. Virol. 76: 392-396 [Abstract] [Full Text]  
  • den Boon, J. A., Chen, J., Ahlquist, P. (2001). Identification of Sequences in Brome Mosaic Virus Replicase Protein 1a That Mediate Association with Endoplasmic Reticulum Membranes. J. Virol. 75: 12370-12381 [Abstract] [Full Text]  
  • Magden, J., Takeda, N., Li, T., Auvinen, P., Ahola, T., Miyamura, T., Merits, A., Kaariainen, L. (2001). Virus-Specific mRNA Capping Enzyme Encoded by Hepatitis E Virus. J. Virol. 75: 6249-6255 [Abstract] [Full Text]  
  • Kujala, P., Ikäheimonen, A., Ehsani, N., Vihinen, H., Auvinen, P., Kääriäinen, L. (2001). Biogenesis of the Semliki Forest Virus RNA Replication Complex. J. Virol. 75: 3873-3884 [Abstract] [Full Text]  
  • Lampio, A., Kilpelainen, I., Pesonen, S., Karhi, K., Auvinen, P., Somerharju, P., Kaariainen, L. (2000). Membrane Binding Mechanism of an RNA Virus-capping Enzyme. J. Biol. Chem. 275: 37853-37859 [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»