Coronavirus Emergence and Evolution: 6+In vitro Analysis of Spike Protein Cleavage of SARS-CoV-2, AcCoV-JC34, and Murine Hepatitis Virus
| dc.contributor.author | Singleton, Deanndria T. | |
| dc.date.accessioned | 2022-08-25T16:19:32Z | |
| dc.date.available | 2022-11-30T07:00:19Z | |
| dc.date.issued | 2022-05 | |
| dc.description.abstract | Members of the Coronaviridae family of viruses represents a highly diverse group that emerges from a wide range of animal reservoirs. Animal reservoirs may vary, from bats and birds to mammals and rodents. Interestingly, some members of Coronaviridae contain a distinct furin cleavage site (FCS) (RXR/KR) at the S1/S2 domain of the spike glycoprotein. The FCS is recognized by the prototypical proprotein convertase, furin, and previous studies show that SARS-CoV-2 contains an FCS, which happens to be atypical in terms of the positioning of the basic (R) residues (RRAR). The FCS can often be found in human, avian and rodent coronaviruses, but are not typically present in SARS-like viruses, including in bat reservoirs. AcCoV-JC34 is a little understood coronavirus (sub-genus Luchacovirus) isolated from Apodemus chevrieri (Chevrier’s field mouse) in Yunnan, China, and is predicted to contain a putative FCS. In this study, first we determined whether the presence of the FCS on AcCoV-JC34 allows for furin cleavage, as it does for SARS-CoV-2. Next, we analyzed the spike protein cleavage of murine hepatitis virus (MHV) strains using furin, and compared our data to prior studies with the goal of understanding disease pathology in mice. To examine these cleavage events, we generated predicted furin cleavage scores using ProP computer software for the S1/S2 domain of selected coronaviruses. We then utilized a fluorogenic peptide cleavage assay in which linear peptide mimics of the S1/S2 domain of selected coronaviruses were mixed with furin protease and buffwe. From the data, we obtained the comparative activity of the protease. We found that although AcCoV-JC34 appears to have an appropriate series of amino acid residues for furin cleavage, our data suggests that it is not cleaved by furin. Further, for MHV, furin cleavage is strain-dependent and can vary across pH values. Overall, this work informs a study of coronavirus evolution, emergence, and pathogenesis with respect to the S protein. | |
| dc.identifier.uri | https://hdl.handle.net/1813/111449 | |
| dc.language.iso | en_US | |
| dc.subject | Biological sciences honors program | |
| dc.subject | Furin Cleavage | |
| dc.subject | Coronaviridae | |
| dc.title | Coronavirus Emergence and Evolution: 6+In vitro Analysis of Spike Protein Cleavage of SARS-CoV-2, AcCoV-JC34, and Murine Hepatitis Virus | |
| dc.type | dissertation or thesis | |
| thesis.degree.discipline | Biological Sciences | |
| thesis.degree.grantor | Cornell University | |
| thesis.degree.level | Bachelor of Science | |
| thesis.degree.name | B.S., Biological Sciences |
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