Understanding the molecular basis of a ubiquitin-dependent kinase and ubiquitin ligase at the interface of inflammation and cell death
<p>Inflammation and programmed cell death (PCD) are seemingly opposing yet intricately linked processes controlled through degradative and non-degradative ubiquitin chains. In this study, I report on two crucial components within apoptosis, a form of PCD, and inflammation. Firstly, the giant i...
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| Other Authors: | |
| Format: | Thesis |
| Language: | English |
| Published: |
2023
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| _version_ | 1826313268183957504 |
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| author | Dietz, L |
| author2 | Elliott, P |
| author_facet | Elliott, P Dietz, L |
| author_sort | Dietz, L |
| collection | OXFORD |
| description | <p>Inflammation and programmed cell death (PCD) are seemingly opposing yet intricately linked processes controlled through degradative and non-degradative
ubiquitin chains. In this study, I report on two crucial components within apoptosis, a form of PCD, and inflammation. Firstly, the giant inhibitor of apoptosis (IAP), BRUCE, and how it mediates caspase inhibition and secondly how the TAK1-TAB complex undergoes activation by Lys63-linked chains.</p>
<p>Here, I present the cryoEM structures of BRUCE alone and in complex with its
regulator SMAC at 3.2 and 3.1 Å resolution, respectively. Complimentary biochemical characterisation shows the capability of BRUCE to ubiquitinate and directly inhibit caspases. This BRUCE-mediated caspase inhibition is released by SMAC, and using structural and biochemical approaches I demonstrate how SMAC utilises a sub nanomolar binding affinity to BRUCE leading to caspase displacement.</p>
<p>Secondly, I present my work on elucidating the structural basis for the TAK1-
TAB complex. Through employing short constructs of TAK1 and TAB2 I show how complex integrity is maintained using biophysical techniques and how Lys6-linked diubiquitin binds to the TAB2 NZF domain using X-ray crystallography. In addition, biochemical assays investigating the activation mechanism of the full-length TAK1 complex suggests longer Lys63-linked ubiquitin chains are required for enhanced activation.</p> |
| first_indexed | 2024-09-25T04:10:24Z |
| format | Thesis |
| id | oxford-uuid:58941660-bd31-4f14-b6be-27d92007646e |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-09-25T04:10:24Z |
| publishDate | 2023 |
| record_format | dspace |
| spelling | oxford-uuid:58941660-bd31-4f14-b6be-27d92007646e2024-06-24T13:06:00ZUnderstanding the molecular basis of a ubiquitin-dependent kinase and ubiquitin ligase at the interface of inflammation and cell deathThesishttp://purl.org/coar/resource_type/c_db06uuid:58941660-bd31-4f14-b6be-27d92007646eBiochemistryEnglishHyrax Deposit2023Dietz, LElliott, P<p>Inflammation and programmed cell death (PCD) are seemingly opposing yet intricately linked processes controlled through degradative and non-degradative ubiquitin chains. In this study, I report on two crucial components within apoptosis, a form of PCD, and inflammation. Firstly, the giant inhibitor of apoptosis (IAP), BRUCE, and how it mediates caspase inhibition and secondly how the TAK1-TAB complex undergoes activation by Lys63-linked chains.</p> <p>Here, I present the cryoEM structures of BRUCE alone and in complex with its regulator SMAC at 3.2 and 3.1 Å resolution, respectively. Complimentary biochemical characterisation shows the capability of BRUCE to ubiquitinate and directly inhibit caspases. This BRUCE-mediated caspase inhibition is released by SMAC, and using structural and biochemical approaches I demonstrate how SMAC utilises a sub nanomolar binding affinity to BRUCE leading to caspase displacement.</p> <p>Secondly, I present my work on elucidating the structural basis for the TAK1- TAB complex. Through employing short constructs of TAK1 and TAB2 I show how complex integrity is maintained using biophysical techniques and how Lys6-linked diubiquitin binds to the TAB2 NZF domain using X-ray crystallography. In addition, biochemical assays investigating the activation mechanism of the full-length TAK1 complex suggests longer Lys63-linked ubiquitin chains are required for enhanced activation.</p> |
| spellingShingle | Biochemistry Dietz, L Understanding the molecular basis of a ubiquitin-dependent kinase and ubiquitin ligase at the interface of inflammation and cell death |
| title | Understanding the molecular basis of a ubiquitin-dependent kinase and ubiquitin ligase at the interface of inflammation and cell death |
| title_full | Understanding the molecular basis of a ubiquitin-dependent kinase and ubiquitin ligase at the interface of inflammation and cell death |
| title_fullStr | Understanding the molecular basis of a ubiquitin-dependent kinase and ubiquitin ligase at the interface of inflammation and cell death |
| title_full_unstemmed | Understanding the molecular basis of a ubiquitin-dependent kinase and ubiquitin ligase at the interface of inflammation and cell death |
| title_short | Understanding the molecular basis of a ubiquitin-dependent kinase and ubiquitin ligase at the interface of inflammation and cell death |
| title_sort | understanding the molecular basis of a ubiquitin dependent kinase and ubiquitin ligase at the interface of inflammation and cell death |
| topic | Biochemistry |
| work_keys_str_mv | AT dietzl understandingthemolecularbasisofaubiquitindependentkinaseandubiquitinligaseattheinterfaceofinflammationandcelldeath |