López, Susana and Yao, Jian-Sheng and Kuhn, Richard J. and Strauss, Ellen G. and Strauss, James H. (1994) Nucleocapsid-glycoprotein interactions required for assembly of alphaviruses. Journal of Virology, 68 (3). pp. 1316-1323. ISSN 0022-538X. http://resolver.caltech.edu/CaltechAUTHORS:LOPjvir94
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We have studied interactions between nucleocapsids and glycoproteins required for budding of alphaviruses, using Ross River virus-Sindbis virus chimeras in which the nucleocapsid protein is derived from one virus and the envelope glycoproteins are derived from the second virus. A virus containing the Ross River virus genome in which the capsid protein had been replaced with that from Sindbis virus was almost nonviable. Nucleocapsids formed in normal numbers in the infected cell, but very little virus was released from the cell. There are 11 amino acid differences between Ross River virus and Sindbis virus in their 33-residue E2 cytoplasmic domains. Site-specific mutagenesis was used to change 9 of these 11 amino acids in the chimera from the Ross River virus to the Sindbis virus sequence in an attempt to adapt the E2 of the chimera to the nucleocapsid. The resulting mutant chimera grew 4 orders of magnitude better than the parental chimeric virus. This finding provides direct evidence for a sequence-specific interaction between the nucleocapsid and the E2 cytoplasmic domain during virus budding. The mutated chimeric virus readily gave rise to large-plaque variants that grew almost as well as Ross River virus, suggesting that additional single amino acid substitutions in the structural proteins can further enhance the interactions between the disparate capsid and the glycoproteins. Unexpectedly, change of E2 residue 394 from lysine (Ross River virus) to glutamic acid (Sindbis virus) was deleterious for the chimera, suggesting that in addition to its role in nucleocapsid-E2 interactions, the N-terminal part of the E2 cytoplasmic domain may be involved in glycoprotein-glycoprotein interactions required to assemble the glycoprotein spikes. The reciprocal chimera, Sindbis virus containing the Ross River virus capsid, also grew poorly. Suppressor mutations arose readily in this chimera, producing a virus that grew moderately well and that formed larger plaques.
|Additional Information:||Copyright © 1994 by the American Society for Microbiology. Received 4 October 1993/Accepted 12 November 1993 S.L. was supported by Public Health Service Fogarty International Research fellowship I FO5 TWO4576-01 JCP(AHR-5) during a sabbatical leave from the Instituto de Biotecnologia/UNAM. This work was supported by NIH grants AI 20612 and AI 10793.|
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|Deposited On:||29 Mar 2006|
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