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 Pugachev, K. V.
Right arrow Articles by Frey, T. K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Pugachev, K. V.
Right arrow Articles by Frey, T. K.

 Previous Article  |  Next Article 

J Virol, January 1998, p. 641-650, Vol. 72, No. 1
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Effects of Defined Mutations in the 5' Nontranslated Region of Rubella Virus Genomic RNA on Virus Viability and Macromolecule Synthesis

Konstantin V. Pugachev and Teryl K. Frey*

Department of Biology, Georgia State University, Atlanta, Georgia 30303

Received 18 July 1997/Accepted 2 October 1997

The 5' end of the genomic RNA of rubella virus (RUB) contains a 14-nucleotide (nt) single-stranded leader (ss-leader) followed by a stem-and-loop structure [5'(+)SL] (nt 15 to 65), the complement of which at the 3' end of the minus-strand RNA [3'(-)SL] has been proposed to function as a promoter for synthesis of genomic plus strands. A second intriguing feature of the 5' end of the RUB genomic RNA is the presence of a short (17 codons) open reading frame (ORF) located between nt 3 and 54; the ORF encoding the viral nonstructural proteins (NSPs) initiates at nt 41 in an alternate translational frame. To address the functional significance of these features, we compared the 5'-terminal sequences of six different strains of RUB, with the result that the short ORF is preserved (although the coding sequence is not conserved) as is the stem part of both the 5'(+)SL and 3'(-)SL, while the upper loop part of both structures varies. Next, using Robo302, an infectious cDNA clone of RUB, we introduced 31 different mutations into the 5'-terminal noncoding region, and their effects on virus replication and macromolecular synthesis were examined. This mutagenesis revealed that the short ORF is not essential for virus replication. The AA dinucleotide at nt 2 and 3 is of critical importance since point mutations and deletions that altered or removed both of these nucleotides were lethal. None of the other mutations within either the ss-leader or the 5'(+)SL [and accordingly within the 3'(-)SL], including deletions of up to 15 nt from the 5'(+)SL and three different multiple-point mutations that lead to destabilization of the 5'(+)SL, were lethal. Some of the mutations within both ss-leader and the 5'(+)SL resulted in viruses that grew to lower titers than the wild-type virus and formed opaque and/or small plaques; in general mutations within the stem had a more profound effect on viral phenotype than did mutations in either the ss-leader or upper loop. Mutations in the 5'(+)SL, but not in the ss-leader, resulted in a significant reduction in NSP synthesis, indicating that this structure is important for efficient translation of the NSP ORF. In contrast, viral plus-strand RNA synthesis was unaffected by the 5'(+)SL mutations as well as the ss-leader mutations, which argues against the proposed function of the 3'(-)SL as a promoter for initiation of the genomic plus-strand RNA.

* Corresponding author. Mailing address: Department of Biology, Georgia State University, 24 Peachtree Center Ave., Atlanta, GA 30303. Phone: (404) 651-3105. Fax: (404) 651-2509. E-mail: biotkf{at}panther.gsu.edu.




This article has been cited by other articles:

  • Claus, C., Tzeng, W.-P., Liebert, U. G., Frey, T. K. (2007). Rubella virus-induced superinfection exclusion studied in cells with persisting replicons. J. Gen. Virol. 88: 2769-2773 [Abstract] [Full Text]  
  • Decca, M. B., Carpio, M. A., Bosc, C., Galiano, M. R., Job, D., Andrieux, A., Hallak, M. E. (2007). Post-translational Arginylation of Calreticulin: A NEW ISOSPECIES OF CALRETICULIN COMPONENT OF STRESS GRANULES. J. Biol. Chem. 282: 8237-8245 [Abstract] [Full Text]  
  • Tzeng, W.-P., Matthews, J. D., Frey, T. K. (2006). Analysis of rubella virus capsid protein-mediated enhancement of replicon replication and mutant rescue.. J. Virol. 80: 3966-3974 [Abstract] [Full Text]  
  • Choi, Y.-J., Yun, S.-I., Kang, S.-Y., Lee, Y.-M. (2006). Identification of 5' and 3' cis-Acting Elements of the Porcine Reproductive and Respiratory Syndrome Virus: Acquisition of Novel 5' AU-Rich Sequences Restored Replication of a 5'-Proximal 7-Nucleotide Deletion Mutant. J. Virol. 80: 723-736 [Abstract] [Full Text]  
  • TORTORICI, M. A., SHAPIRO, B. A., PATTON, J. T. (2006). A base-specific recognition signal in the 5' consensus sequence of rotavirus plus-strand RNAs promotes replication of the double-stranded RNA genome segments. RNA 12: 133-146 [Abstract] [Full Text]  
  • Timchenko, N. A., Wang, G.-L., Timchenko, L. T. (2005). RNA CUG-binding Protein 1 Increases Translation of 20-kDa Isoform of CCAAT/Enhancer-binding Protein {beta} by Interacting with the {alpha} and {beta} Subunits of Eukaryotic Initiation Translation Factor 2. J. Biol. Chem. 280: 20549-20557 [Abstract] [Full Text]  
  • Gorchakov, R., Hardy, R., Rice, C. M., Frolov, I. (2004). Selection of Functional 5' cis-Acting Elements Promoting Efficient Sindbis Virus Genome Replication. J. Virol. 78: 61-75 [Abstract] [Full Text]  
  • Timchenko, L. T., Iakova, P., Welm, A. L., Cai, Z.-J., Timchenko, N. A. (2002). Calreticulin Interacts with C/EBP{alpha} and C/EBP{beta} mRNAs and Represses Translation of C/EBP Proteins. Mol. Cell. Biol. 22: 7242-7257 [Abstract] [Full Text]  
  • Lee, J.-Y., Bowden, D. S. (2000). Rubella Virus Replication and Links to Teratogenicity. Clin. Microbiol. Rev. 13: 571-587 [Abstract] [Full Text]  
  • Lund, K. D., Chantler, J. K. (2000). Mapping of Genetic Determinants of Rubella Virus Associated with Growth in Joint Tissue. J. Virol. 74: 796-804 [Abstract] [Full Text]  
  • Chen, M.-H., Frey, T. K. (1999). Mutagenic Analysis of the 3' cis-Acting Elements of the Rubella Virus Genome. J. Virol. 73: 3386-3403 [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»