The rhabdovirus vesicular stomatitis virus (VSV) is a model system for the study of the group of viruses possessing non-segmented single- stranded RNA genomes of the negative strand sense. It includes such human pathogens as measles, mumps, rabies, respiratory syncytial virus, human parainfluenza viruses, and many viruses that infect economically important animals. A central question concerning the growth of these viruses is: by what mechanism do they control the expression of their genetic information through transcription and replication of the genome. The goals of the research in this application are two fold. First, we will continue our study of the form and function of the proteins involved in the assembly of newly replicating nucleocapsids. We will attempt to clarify the relationship between the soluble forms of the N protein in the cell and the various RNA-binding activities of the molecule (encapsidation of genome-length RNA, encapsidation of free leader RNA, and binding to mRNAs). Secondly, we will perform a molecular genetic analysis of cis-acting regulatory sequences in controlling the processes of transcription, replication, and nucleocapsid assembly. We will do this by producing a full-length cDNA copy of the genome RNA of a defective-interfering (DI) particle of the virus, and placing this cDNA under the control of a bacterial promoter in the plasmid pPMI. In this way, we will be able to produce RNA of identical sequence to the DI genome in vitro. We have developed a system to assemble the RNA into functional nucleocapsids in vitro, and will use this system in conjunction with site-directed mutagenesis of the cloned DI genome to assess the role of specific RNA sequences in nucleocapsid assembly, initiation and termination of transcription, intragenic pausing of the virion polymerase, and to address questions relating to the mechanism of interference of standard virus by DI particles. We will also investigate the possible construction and use of chimeric DI nucleocapsids as vectors for gene expression in mammalian cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI022116-05
Application #
3132823
Study Section
Experimental Virology Study Section (EVR)
Project Start
1986-06-01
Project End
1994-05-31
Budget Start
1990-06-01
Budget End
1991-05-31
Support Year
5
Fiscal Year
1990
Total Cost
Indirect Cost
Name
Mount Sinai School of Medicine
Department
Type
Schools of Medicine
DUNS #
City
New York
State
NY
Country
United States
Zip Code
10029
Richardson, J C; Peluso, R W (1996) Inhibition of VSV genome RNA replication but not transcription by monoclonal antibodies specific for the viral P protein. Virology 216:26-34
Moscona, A; Peluso, R W (1996) Analysis of human parainfluenza virus 3 receptor binding variants: evidence for the use of a specific sialic acid-containing receptor. Microb Pathog 20:179-84
Kim, S H; Chung, Y B (1996) Isolation of a mutant bacteriophage T7 deleted in nonessential genetic elements, gene 19.5 and m. Virology 216:20-5
Peluso, R W; Richardson, J C; Talon, J et al. (1996) Identification of a set of proteins (C' and C) encoded by the bicistronic P gene of the Indiana serotype of vesicular stomatitis virus and analysis of their effect on transcription by the viral RNA polymerase. Virology 218:335-42
Huberman, K; Peluso, R W; Moscona, A (1995) Hemagglutinin-neuraminidase of human parainfluenza 3: role of the neuraminidase in the viral life cycle. Virology 214:294-300
Moscona, A; Peluso, R W (1993) Relative affinity of the human parainfluenza virus type 3 hemagglutinin-neuraminidase for sialic acid correlates with virus-induced fusion activity. J Virol 67:6463-8
Moscona, A; Peluso, R W (1993) Persistent infection with human parainfluenza virus 3 in CV-1 cells: analysis of the role of defective interfering particles. Virology 194:399-402
Moscona, A; Peluso, R W (1992) Fusion properties of cells infected with human parainfluenza virus type 3: receptor requirements for viral spread and virus-mediated membrane fusion. J Virol 66:6280-7
La Ferla, F M; Peluso, R W (1989) The 1:1 N-NS protein complex of vesicular stomatitis virus is essential for efficient genome replication. J Virol 63:3852-7
Peluso, R W (1988) Kinetic, quantitative, and functional analysis of multiple forms of the vesicular stomatitis virus nucleocapsid protein in infected cells. J Virol 62:2799-807

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