The non-segmented negative-strand RNA viruses comprise some of the most serious and uncontrolled human viral pathogens. Against such a group of pathogens there are two effective research strategies: to address each virus individually and develop virus-specific therapeutics or prophylactics, or to select a prototype virus and examine every aspect of its replication cycle in depth, with the goal of developing general concepts for intervention. Both approaches are valid, but the latter generally has more far-reaching long- term consequences. Among the non-segmented negative-strand RNA viruses, the rhabdovirus vesicular stomatitis virus (VSV) has been studied as the prototype for several decades, with the result that it is one of the best-understood animal viruses. The development of methods to engineer changes into the genome of negative-strand RNA viruses at the cDNA level and recover infectious virus revolutionized research on these viruses. It is now possible to investigate the effect of specific mutations on every aspect of the biology of negative-strand RNA viruses. In the past grant period major advances were made in defining the cis-acting signals involved in the processes of RNA transcription and replication. Our goal now is to determine the mechanisms by which these cis-acting elements control transcription and replication, to determine how they interact with trans-acting factors, and to analyze their importance in the viral life cycle. We propose to do this by addressing the following specific aims: 1. To identify residues involved in nucleocapsid protein (N):RNA encapsidation by structure-guided mutagenesis using semi-randomized oligonucleotides introduced into an infectious VSV cDNA and allowing natural selection to determine the range of possible alternative genotypes and phenotypes. 2. To identify residues critical for N protein flexibility and N:N interactions using structure-guided mutations. 3. To investigate the RNA sequence requirements for the encapsidation of specific RNAs. 4. To use the power of natural selection to identify N protein:RNA interacting residues. 5. To analyze the mechanism of polymerase attenuation at gene junctions. 6. To test a model for the mechanism of transcriptional attenuation.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Research Project (R01)
Project #
Application #
Study Section
Virology - B Study Section (VIRB)
Program Officer
Cassetti, Cristina
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Virginia
Schools of Medicine
United States
Zip Code
Mondal, Arindam; Victor, Ken G; Pudupakam, R S et al. (2014) Newly identified phosphorylation site in the vesicular stomatitis virus P protein is required for viral RNA synthesis. J Virol 88:1461-72
Rainsford, Edward W; Harouaka, Djamila; Wertz, Gail W (2010) Importance of hydrogen bond contacts between the N protein and RNA genome of vesicular stomatitis virus in encapsidation and RNA synthesis. J Virol 84:1741-51
Galloway, Summer E; Wertz, Gail W (2009) A temperature sensitive VSV identifies L protein residues that affect transcription but not replication. Virology 388:286-93
Harouaka, Djamila; Wertz, Gail W (2009) Mutations in the C-terminal loop of the nucleocapsid protein affect vesicular stomatitis virus RNA replication and transcription differentially. J Virol 83:11429-39
Galloway, Summer E; Wertz, Gail W (2008) S-adenosyl homocysteine-induced hyperpolyadenylation of vesicular stomatitis virus mRNA requires the methyltransferase activity of L protein. J Virol 82:12280-90
Barr, John N (2007) Bunyavirus mRNA synthesis is coupled to translation to prevent premature transcription termination. RNA 13:731-6
Flanagan, E Brian; Schoeb, Trenton R; Wertz, Gail W (2003) Vesicular stomatitis viruses with rearranged genomes have altered invasiveness and neuropathogenesis in mice. J Virol 77:5740-8
Hardy, R W; Wertz, G W (2000) The Cys(3)-His(1) motif of the respiratory syncytial virus M2-1 protein is essential for protein function. J Virol 74:5880-5
Whelan, S P; Barr, J N; Wertz, G W (2000) Identification of a minimal size requirement for termination of vesicular stomatitis virus mRNA: implications for the mechanism of transcription. J Virol 74:8268-76
Whelan, S P; Wertz, G W (1999) Regulation of RNA synthesis by the genomic termini of vesicular stomatitis virus: identification of distinct sequences essential for transcription but not replication. J Virol 73:297-306

Showing the most recent 10 out of 31 publications