Structural and cellular biology of Teneurins and other neuronal cell adhesion receptors
<p>The development of the nervous system is a complicated process that rests on correct cell migration and cell-to-cell (<em>trans</em>) communication. These in turn rely on cell adhesion molecules which perform most of the interactions happening during these processes. Traditional...
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| Format: | Thesis |
| Language: | English |
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2023
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| _version_ | 1826314916836933632 |
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| author | Berbeira Santana, M |
| author2 | Seiradake, E |
| author_facet | Seiradake, E Berbeira Santana, M |
| author_sort | Berbeira Santana, M |
| collection | OXFORD |
| description | <p>The development of the nervous system is a complicated process that rests on correct cell migration and cell-to-cell (<em>trans</em>) communication. These in turn rely on cell adhesion molecules which perform most of the interactions happening during these processes. Traditionally, the roles of these proteins have been studied <em>in vivo</em> using knock-outs and similar strategies, which delete whole proteins and all their subsequent functions. Also, they do not provide a mechanistic insight into how the interactions happen. To overcome this, structural biology coupled with genetic engineering and cell biology posits as a solution.</p>
<p>In this thesis, the main aim was to understand the role of Teneurin <em>trans</em> homophilic interactions using cryogenic electron microscopy (cryoEM) and cell biology. Dimeric structures of all four mTeneurin2 splice site variants show the same binding interface, which overlaps with that of Latrophilin. Structure-based mutants allow for binding to only one of the two proteins. Also, using aggregation assays I showed the interplay between the Teneurin-Teneurin and Teneurin-Latrophilin interactions.</p>
<p>In addition, in this thesis a potential oligomeric cryoEM map of mTeneurin4 A1B0 is presented. However, several issues during sample preparation, data acquisition and data processing hindered the full interpretation of the map. Several solutions were tested, but none provided a map of sufficient quality to place models. Also, two monomeric maps of two splice site variants of mTeneurin3 are presented in this thesis, alongside the monomeric maps for both fly Teneurins (Teneurin-m and -a). Moreover, a preliminary low-resolution map shows a novel dimeric conformation of Teneurin-a.</p>
<p>Finally, using different cell biology techniques, such as cell-based aggregation assays and cell binding assays, different tools (nanobodies and structure-based mutants) to study the GPC3- Unc5D interaction were validated. In addition, a cis-masking effect between the <em>cis</em> and <em>trans</em> GPC3-Unc5D interaction was discovered.</p> |
| first_indexed | 2024-12-09T03:14:53Z |
| format | Thesis |
| id | oxford-uuid:64abb59a-f62b-414f-bfe3-0a931586bda1 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-12-09T03:14:53Z |
| publishDate | 2023 |
| record_format | dspace |
| spelling | oxford-uuid:64abb59a-f62b-414f-bfe3-0a931586bda12024-10-16T15:06:27ZStructural and cellular biology of Teneurins and other neuronal cell adhesion receptorsThesishttp://purl.org/coar/resource_type/c_db06uuid:64abb59a-f62b-414f-bfe3-0a931586bda1Structural biologyCell biologyEnglishHyrax Deposit2023Berbeira Santana, MSeiradake, EZitzmann, N<p>The development of the nervous system is a complicated process that rests on correct cell migration and cell-to-cell (<em>trans</em>) communication. These in turn rely on cell adhesion molecules which perform most of the interactions happening during these processes. Traditionally, the roles of these proteins have been studied <em>in vivo</em> using knock-outs and similar strategies, which delete whole proteins and all their subsequent functions. Also, they do not provide a mechanistic insight into how the interactions happen. To overcome this, structural biology coupled with genetic engineering and cell biology posits as a solution.</p> <p>In this thesis, the main aim was to understand the role of Teneurin <em>trans</em> homophilic interactions using cryogenic electron microscopy (cryoEM) and cell biology. Dimeric structures of all four mTeneurin2 splice site variants show the same binding interface, which overlaps with that of Latrophilin. Structure-based mutants allow for binding to only one of the two proteins. Also, using aggregation assays I showed the interplay between the Teneurin-Teneurin and Teneurin-Latrophilin interactions.</p> <p>In addition, in this thesis a potential oligomeric cryoEM map of mTeneurin4 A1B0 is presented. However, several issues during sample preparation, data acquisition and data processing hindered the full interpretation of the map. Several solutions were tested, but none provided a map of sufficient quality to place models. Also, two monomeric maps of two splice site variants of mTeneurin3 are presented in this thesis, alongside the monomeric maps for both fly Teneurins (Teneurin-m and -a). Moreover, a preliminary low-resolution map shows a novel dimeric conformation of Teneurin-a.</p> <p>Finally, using different cell biology techniques, such as cell-based aggregation assays and cell binding assays, different tools (nanobodies and structure-based mutants) to study the GPC3- Unc5D interaction were validated. In addition, a cis-masking effect between the <em>cis</em> and <em>trans</em> GPC3-Unc5D interaction was discovered.</p> |
| spellingShingle | Structural biology Cell biology Berbeira Santana, M Structural and cellular biology of Teneurins and other neuronal cell adhesion receptors |
| title | Structural and cellular biology of Teneurins and other neuronal cell adhesion receptors |
| title_full | Structural and cellular biology of Teneurins and other neuronal cell adhesion receptors |
| title_fullStr | Structural and cellular biology of Teneurins and other neuronal cell adhesion receptors |
| title_full_unstemmed | Structural and cellular biology of Teneurins and other neuronal cell adhesion receptors |
| title_short | Structural and cellular biology of Teneurins and other neuronal cell adhesion receptors |
| title_sort | structural and cellular biology of teneurins and other neuronal cell adhesion receptors |
| topic | Structural biology Cell biology |
| work_keys_str_mv | AT berbeirasantanam structuralandcellularbiologyofteneurinsandotherneuronalcelladhesionreceptors |