Type of Document Master's Thesis Author Elbahesh, Husni Author's Email Address husni_elbahesh@hotmail.com URN etd-11222005-153855 Title Functional Analysis of the Murine Oligoadenylate Synthetase 1b (Oas1b) Degree Master of Science Department Biology Advisory Committee
Advisor Name Title Margo A. Brinton Committee Chair Teryl K. Frey Committee Member Zehava Eichenbaum Committee Member Keywords
- West Nile virus
- viral RNA
- Oligoadenylate Synthetase
- Flavivirus
Date of Defense 2005-08-17 Availability unrestricted Abstract The flavivirus resistance gene, Flv, in mice has been identified as 2'-5' oligoadenylate synthetase 1b (Oas1b). Susceptible mice produce a protein that is truncated (Oas1btr) at the C-terminus due to a premature stop codon encoded by a C820T transition. Mice produce 8 Oas1 proteins, Oas1a-Oas1h. In the present study, Oas1a, Oas1b and Oas1btr were expressed as MBP-fusion proteins in bacteria and purified. 2-5A synthetase activity was demonstrated using MBP-Oas1a, while neither MBP-Oas1b nor MBP-Oas1btr were functionally active. The 2-5A synthetase activity of MBP-Oas1a was inhibited in a dose-dependent manner by the addition of MBP-Oas1b but not MBPOas1btr. Finally, three RNA probes were synthesized from the 3' end of the WNV Eg101 genome and used to test the ability of the expressed Oas1 proteins to bind to viral RNA. Results of the RNA binding activity assays suggest Oas1 proteins may specifically interact with regions of WNV RNA.Files
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