Abstract

The overall aim of the proposed research program is the identify the functions of the proteins encoded by reovirus.
Six specific aims are proposed. The first is to isolate the native forms of all reovirus proteins from cells infected with a powerful mammalian expression vector into which the reovirus genome segments have been cloned. The isolated proteins will be examined for enzymic activities and ability to bind to RNA and other proteins, both viral and cellular. Second, transient expression vectors will be used to examine the effect of reovirus proteins on DNA replication, transcription and protein synthesis. Genetically engineered genome segments will be used to identify functional domains. The intracellular location of reovirus proteins into which reovirus RNA species have been lipofected singly and in recombinant vaccinia virus strains containing the various genome segments. Third, techniques will be developed to introduce genetically engineered genome segments into reovirus genomes so as to permit study of the effect of alterations in functional domains on viral phenotype. Fourth, temperature-sensitive mutants and anti-sense technology will be used to determine at what stage inability of individual viral proteins to function arrests virus multiplication, thereby providing further clues to viral protein function. Fifth, selected SORFs in reovirus serotype 1, 2 and 3 cell receptors will be identified in gene libraries and sequenced in order to identify the nature of the receptors.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI008909-23
Application #
3124480
Study Section
Virology Study Section (VR)
Project Start
1974-09-01
Project End
1995-03-31
Budget Start
1991-04-01
Budget End
1992-03-31
Support Year
23
Fiscal Year
1991
Total Cost
Indirect Cost

  Institution

Name
Duke University
Department
Type
Schools of Medicine
DUNS #
071723621
City
Durham
State
NC
Country
United States
Zip Code
27705

  Publications

Joklik, W K (1998) Assembly of the reovirus genome. Curr Top Microbiol Immunol 233:57-68
Roner, M R; Nepliouev, I; Sherry, B et al. (1997) Construction and characterization of a reovirus double temperature-sensitive mutant. Proc Natl Acad Sci U S A 94:6826-30
Joklik, W K; Roner, M R (1996) Molecular recognition in the assembly of the segmented reovirus genome. Prog Nucleic Acid Res Mol Biol 53:249-81
Roner, M R; Lin, P N; Nepluev, I et al. (1995) Identification of signals required for the insertion of heterologous genome segments into the reovirus genome. Proc Natl Acad Sci U S A 92:12362-6
Joklik, W K; Roner, M R (1995) What reassorts when reovirus genome segments reassort? J Biol Chem 270:4181-4
Larson, S M; Antczak, J B; Joklik, W K (1994) Reovirus exists in the form of 13 particle species that differ in their content of protein sigma 1. Virology 201:303-11
Starnes, M C; Joklik, W K (1993) Reovirus protein lambda 3 is a poly(C)-dependent poly(G) polymerase. Virology 193:356-66
Roner, M R; Roner, L A; Joklik, W K (1993) Translation of reovirus RNA species m1 can initiate at either of the first two in-frame initiation codons. Proc Natl Acad Sci U S A 90:8947-51
Xu, P; Miller, S E; Joklik, W K (1993) Generation of reovirus core-like particles in cells infected with hybrid vaccinia viruses that express genome segments L1, L2, L3, and S2. Virology 197:726-31
Antczak, J B; Joklik, W K (1992) Reovirus genome segment assortment into progeny genomes studied by the use of monoclonal antibodies directed against reovirus proteins. Virology 187:760-76

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