Functional characterization of oligoadenylate synthetase (OAS) by dual polymerase-endoribonuclease assay

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dc.contributor.authorLubna, Nikhat
dc.contributor.examiningcommitteePortet, Stephanie (Mathematics)en_US
dc.contributor.examiningcommitteeKhajehpour, Mazdak (Chemistry)en_US
dc.contributor.supervisorMcKenna, Sean
dc.date.accessioned2023-03-14T19:10:38Z
dc.date.available2023-03-14T19:10:38Z
dc.date.copyright2023-02-28
dc.date.issued2023-02-28
dc.date.submitted2023-02-28T20:28:17Zen_US
dc.degree.disciplineChemistryen_US
dc.degree.levelMaster of Science (M.Sc.)en_US
dc.description.abstractThe innate immune system includes a class of viral double stranded RNA (dsRNA) binding enzymes known as 2’-5’-oligoadenylate synthetases (OAS). The OAS family detects viral dsRNA and initiates downstream processes. The binding of viral dsRNA by OAS enables catalysis of substrate ATP into 2′-5′-linked oligoadenylate chains (2-5A). 2-5A chains longer than 3 nucleotides then activate an RNA degrading enzyme named RNase L, which in turn non-specifically degrades cellular and viral RNA causing host cell death. My research group has been investigating the interaction between the smallest OAS family member, OAS1 (42 kDa), with a double-stranded region of the West Nile virus (WNV) RNA genome. This region of the genome is conserved amongst the Flaviviridae family of viruses, of which WNV is a member. Therefore, the goal of my work is to see whether similar RNA regions in other Flaviviridae members using Zika Virus, Japanese Encephalitis Virus, WNV and Dengue Virus as model systems for studying this interaction. The approach will be to produce viral RNA from the conserved regions and determine the minimal RNA required for binding of OAS1 and catalytic activation of OAS1. I have produced a dual OAS-RNase L activity assay which provides data in shorter time intervals, measures OAS activity and RNase L activity simultaneously, and requires very low concentrations of reactants. The data obtained from this assay will be used to assess activation of OAS enzyme by flavivirus RNA and help paint a more complete picture of OAS mechanism of action.en_US
dc.description.noteMay 2023en_US
dc.identifier.urihttp://hdl.handle.net/1993/37205
dc.language.isoengen_US
dc.rightsopen accessen_US
dc.subjectoligoadenylateen_US
dc.subjectsynthetaseen_US
dc.subjectRNaseLen_US
dc.subjectendoribonucleaseen_US
dc.subjectassayen_US
dc.subjectFlavivirusen_US
dc.subjectJapaneseen_US
dc.subjectencephalitisen_US
dc.subjectDengueen_US
dc.subjectZikaen_US
dc.subjectInnateen_US
dc.subjectImmuneen_US
dc.subjectenzymeen_US
dc.subjectmechanismen_US
dc.subjectPRRen_US
dc.subjectPAMPen_US
dc.subjectOASen_US
dc.subjectNikhaten_US
dc.subjectLubnaen_US
dc.subjectAmiten_US
dc.subjectKoulen_US
dc.subjectSeanen_US
dc.subjectMcKennaen_US
dc.titleFunctional characterization of oligoadenylate synthetase (OAS) by dual polymerase-endoribonuclease assayen_US
dc.typemaster thesisen_US
local.subject.manitobanoen_US
oaire.awardNumberRGPIN-2015-06142en_US
oaire.awardTitleDiscovery Grants Program - Individualen_US
oaire.awardURIhttps://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=730026en_US
project.funder.identifierhttps://doi.org/10.13039/501100000038en_US
project.funder.nameNatural Sciences and Engineering Research Council of Canadaen_US
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