Attempts will be made to answer several questions concerning the nature of the functions encoded in the reovirus genome and concerning reovirus multiplication. They are: a. Reovirus morphogenesis will be estudied in vivo by analyzing the nature of the RNA and protein components of progressively more mature immature reovirus particles, starting with the first complexes into which the ten individual species of ssRNA molecules are assembled. It will also be studied in vitro by reacting various combinations of native reovirus proteins with ssRNA molecules and isolating and characterizing the complexes that result. b. Native reovirus proteins will be examined for possession of the five enzyme activities that are exhibited by reovirus cores. Such proteins will be isolated from bacteria transformed by expression vectors into which reovirus genes have been cloned. c. Attempts will be made to elucidate the functions of reovirus proteins by examining in detail the multiplication cycles of ts mutants at nonpermissive temperatures. Mutants with lesions in each of the ten genes are available. Multiplication cycles will be analyzed in terms of the kinetics of synthesis of ss and dsRNA species and reovirus proteins, as well as of the various particles on the pathway of reovirus morphogenesis. d. The various reovirus messenger RNA species are not translated with equal efficiency. The importance of regions upstream from and around initiation codons as elements that control efficiency of translation will e investigated by introducing mutations, deletions and insertions into them. be. The evolutionary and functional relationships of the three reovirus serotypes will be studied by sequencing cognate sets of their genes and examining cognate sets of their proteins for similarity of tertiary structure and for the nature of their antigenic and functional domains. f. The mode of the anti-reovirus activity of ribavirin, and various pure interferon species will be examined by determining which reactions of the reovirus multiplication cycle they inhibit. Hopefully, these results can be extended and applied to infections by rotaviruses and orbiviruses of medical importance.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI008909-20
Application #
3124478
Study Section
Virology Study Section (VR)
Project Start
1974-09-01
Project End
1989-08-31
Budget Start
1987-09-01
Budget End
1988-08-31
Support Year
20
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Duke University
Department
Type
Schools of Medicine
DUNS #
071723621
City
Durham
State
NC
Country
United States
Zip Code
27705
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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