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 Tubulekas, I.
Right arrow Articles by Liljeström, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Tubulekas, I.
Right arrow Articles by Liljeström, P.

 Previous Article  |  Next Article 

J Virol, April 1998, p. 2825-2831, Vol. 72, No. 4
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Suppressors of Cleavage-Site Mutations in the p62 Envelope Protein of Semliki Forest Virus Reveal Dynamics in Spike Structure and Function

Ioannis Tubulekas1 and Peter Liljeström2,3,*

Department of Biosciences at Novum, Karolinska Institute, Huddinge,1 and Microbiology and Tumorbiology Center, Karolinska Institute,2 and Department of Vaccine Research, Swedish Institute for Infectious Disease Control,3 Stockholm, Sweden

Received 6 October 1997/Accepted 17 December 1997

The E2 spike glycoprotein of Semliki Forest virus is produced as a p62 precursor protein, which is cleaved by host proteases to its mature form, E2. Cleavage is not necessary for particle formation or release but is necessary for infectivity. Previous results had shown that phenotypic revertants of cleavage-deficient p62 mutants are generated, and here we show that these may contain second-site suppressor mutations in the vicinity of the cleavage site. These hot-spot sites were mutated to abolish the generation of such suppressor mutations; however, secondary mutations in another distant domain of the E2 protein appeared instead, all of which still caused cleavage-deficient mutations. Such mutants grew very poorly and were inefficient in virus entry and release. The mutated sites define domains of the spike protein which probably interact to regulate its structure and function. Because of their highly attenuated phenotype and the lower probability of reversion, the new mutations close to the cleavage site were used to make new helper vectors for packaging of recombinant RNA into infectious particles, thus increasing further the biosafety of the vector system based on the Semliki Forest virus replicon.

* Corresponding author. Mailing address: Microbiology and Tumorbiology Center, Karolinska Institute, Box 280, S-171 77 Stockholm, Sweden. Phone: 46-8-728 6306. Fax: 46-8-319 587. E-mail: Peter.Liljestrom{at}mtc.ki.se.




This article has been cited by other articles:

  • Wu, S.-R., Haag, L., Sjoberg, M., Garoff, H., Hammar, L. (2008). The Dynamic Envelope of a Fusion Class II Virus: E3 DOMAIN OF GLYCOPROTEIN E2 PRECURSOR IN SEMLIKI FOREST VIRUS PROVIDES A UNIQUE CONTACT WITH THE FUSION PROTEIN E1. J. Biol. Chem. 283: 26452-26460 [Abstract] [Full Text]  
  • Elshuber, S., Mandl, C. W. (2005). Resuscitating Mutations in a Furin Cleavage-Deficient Mutant of the Flavivirus Tick-Borne Encephalitis Virus. J. Virol. 79: 11813-11823 [Abstract] [Full Text]  
  • Hidmark, A. S., McInerney, G. M., Nordstrom, E. K. L., Douagi, I., Werner, K. M., Liljestrom, P., Hedestam, G. B. K. (2005). Early Alpha/Beta Interferon Production by Myeloid Dendritic Cells in Response to UV-Inactivated Virus Requires Viral Entry and Interferon Regulatory Factor 3 but Not MyD88. J. Virol. 79: 10376-10385 [Abstract] [Full Text]  
  • Sjoberg, M., Garoff, H. (2003). Interactions between the Transmembrane Segments of the Alphavirus E1 and E2 Proteins Play a Role in Virus Budding and Fusion. J. Virol. 77: 3441-3450 [Abstract] [Full Text]  
  • Hammar, L., Markarian, S., Haag, L., Lankinen, H., Salmi, A., Cheng, R. H. (2003). Prefusion Rearrangements Resulting in Fusion Peptide Exposure in Semliki Forest Virus. J. Biol. Chem. 278: 7189-7198 [Abstract] [Full Text]  
  • Smit, J. M., Klimstra, W. B., Ryman, K. D., Bittman, R., Johnston, R. E., Wilschut, J. (2001). PE2 Cleavage Mutants of Sindbis Virus: Correlation between Viral Infectivity and pH-Dependent Membrane Fusion Activation of the Spike Heterodimer. J. Virol. 75: 11196-11204 [Abstract] [Full Text]  
  • Boehme, K. W., Williams, J. C., Johnston, R. E., Heidner, H. W. (2000). Linkage of an alphavirus host-range restriction to the carbohydrate-processing phenotypes of the host cell. J. Gen. Virol. 81: 161-170 [Abstract] [Full Text]  
  • Smerdou, C., Liljeström, P. (1999). Two-Helper RNA System for Production of Recombinant Semliki Forest Virus Particles. J. Virol. 73: 1092-1098 [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»