Integrative structural biology of viruses
<p>Picornaviruses include significant infectious agents and are challenging structural targets owing to their pathogenicity. They share a common architecture, but subtle differences in their structure mediate tissue tropism, receptor binding and uncoating cues. This thesis focuses on members o...
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| Format: | Thesis |
| Language: | English |
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2019
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| _version_ | 1797100118924591104 |
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| author | Duyvesteyn, HME |
| author2 | Huiskonen, J |
| author_facet | Huiskonen, J Duyvesteyn, HME |
| author_sort | Duyvesteyn, HME |
| collection | OXFORD |
| description | <p>Picornaviruses include significant infectious agents and are challenging structural targets owing to their pathogenicity. They share a common architecture, but subtle differences in their structure mediate tissue tropism, receptor binding and uncoating cues. This thesis focuses on members of the Enterovirus and Aphthovirus genera, and aims to contribute structural understanding of their lifecycle and the forces that underpin and modulate capsid stability to ultimately inspire novel therapeutics and vaccines. I take advantage of improvements in cryoEM which enable structural biologists to routinely solve complex structures at close to atomic resolution and resolve sample heterogeneity. </p>
<p>EV-F3 is a non-pathogenic enterovirus. Presented crystallographic structures or EV-F3 highlight a potentially druggable site, and soaks with siallactose suggest a cellular receptor. Cryo-EM investigations show intermediate states, confirming that heating can cause transitions to altered particle states seen in other enteroviruses. Long-wavelength X-ray data indicate the presence of K<sup>+</sup>, but not Ca<sup>2+</sup> ions within the capsid.</p>
<p>I present structural characterisation of recombinant VLP vaccine candidates for poliovirus (PV) and foot-and-mouth disease virus (FMDV). For FMDV, I show a continuum of cryo-EM structures with a potential S-S bridge at the 2-fold interface and varied 2-fold structures. A pentamer subassembly is characterised and analysis of metal ion composition in FMDV VLPs gives no evidence for bound Ca<sup>2+</sup> ions. </p>
<p>Single-particle cryo-EM experiments of yeast-derived thermostabilised PV-1 and PV-3 VLPs show both C and D antigenic states to be present for PV-1 and demonstrate a predominance of D-antigenic states for PV-3. </p>
<p>Finally, preliminary investigations into aphthovirus uncoating include attempts to express full length α<sub>v</sub>β<sub>6</sub> integrin receptor and a preliminary structure of FMDV bound to α<sub>v</sub>β<sub>3</sub> head-group, demonstrating VLP affinity for virus receptors. I demonstrate the potential of caged protons for time-resolved structural studies of FMDV uncoating and present a novel expanded particle structure of an aphthovirus.</p> |
| first_indexed | 2024-03-07T05:33:13Z |
| format | Thesis |
| id | oxford-uuid:e2fd4ad8-6060-436d-8f54-56547f257e1d |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T05:33:13Z |
| publishDate | 2019 |
| record_format | dspace |
| spelling | oxford-uuid:e2fd4ad8-6060-436d-8f54-56547f257e1d2022-03-27T10:05:38ZIntegrative structural biology of virusesThesishttp://purl.org/coar/resource_type/c_db06uuid:e2fd4ad8-6060-436d-8f54-56547f257e1dEnterovirusesStructural biologyPicornavirusesVirus-like particlesAphthovirusesFoot-and-mouth disease virusTransmission electron microscopyX-ray crystallographyPoliovirusVirologyEnglishHyrax Deposit2019Duyvesteyn, HMEHuiskonen, JStuart, DOrville, AGrimes, JHogle, J<p>Picornaviruses include significant infectious agents and are challenging structural targets owing to their pathogenicity. They share a common architecture, but subtle differences in their structure mediate tissue tropism, receptor binding and uncoating cues. This thesis focuses on members of the Enterovirus and Aphthovirus genera, and aims to contribute structural understanding of their lifecycle and the forces that underpin and modulate capsid stability to ultimately inspire novel therapeutics and vaccines. I take advantage of improvements in cryoEM which enable structural biologists to routinely solve complex structures at close to atomic resolution and resolve sample heterogeneity. </p> <p>EV-F3 is a non-pathogenic enterovirus. Presented crystallographic structures or EV-F3 highlight a potentially druggable site, and soaks with siallactose suggest a cellular receptor. Cryo-EM investigations show intermediate states, confirming that heating can cause transitions to altered particle states seen in other enteroviruses. Long-wavelength X-ray data indicate the presence of K<sup>+</sup>, but not Ca<sup>2+</sup> ions within the capsid.</p> <p>I present structural characterisation of recombinant VLP vaccine candidates for poliovirus (PV) and foot-and-mouth disease virus (FMDV). For FMDV, I show a continuum of cryo-EM structures with a potential S-S bridge at the 2-fold interface and varied 2-fold structures. A pentamer subassembly is characterised and analysis of metal ion composition in FMDV VLPs gives no evidence for bound Ca<sup>2+</sup> ions. </p> <p>Single-particle cryo-EM experiments of yeast-derived thermostabilised PV-1 and PV-3 VLPs show both C and D antigenic states to be present for PV-1 and demonstrate a predominance of D-antigenic states for PV-3. </p> <p>Finally, preliminary investigations into aphthovirus uncoating include attempts to express full length α<sub>v</sub>β<sub>6</sub> integrin receptor and a preliminary structure of FMDV bound to α<sub>v</sub>β<sub>3</sub> head-group, demonstrating VLP affinity for virus receptors. I demonstrate the potential of caged protons for time-resolved structural studies of FMDV uncoating and present a novel expanded particle structure of an aphthovirus.</p> |
| spellingShingle | Enteroviruses Structural biology Picornaviruses Virus-like particles Aphthoviruses Foot-and-mouth disease virus Transmission electron microscopy X-ray crystallography Poliovirus Virology Duyvesteyn, HME Integrative structural biology of viruses |
| title | Integrative structural biology of viruses |
| title_full | Integrative structural biology of viruses |
| title_fullStr | Integrative structural biology of viruses |
| title_full_unstemmed | Integrative structural biology of viruses |
| title_short | Integrative structural biology of viruses |
| title_sort | integrative structural biology of viruses |
| topic | Enteroviruses Structural biology Picornaviruses Virus-like particles Aphthoviruses Foot-and-mouth disease virus Transmission electron microscopy X-ray crystallography Poliovirus Virology |
| work_keys_str_mv | AT duyvesteynhme integrativestructuralbiologyofviruses |