Capsaspora owczarzaki as a unicellular model to study the pre-metazoan integrin adhesome
[eng] Animal multicellular structure depends on three general mechanisms: cell adhesion, cell communication and cell differentiation. Some of these mechanisms were thought to be specific to animals. Genome sequencing of unicellular organisms that are closely related to animals has revealed that the...
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| Tipo de recurso: | tesis doctoral |
| Estado: | Versión publicada |
| Fecha de publicación: | 2019 |
| País: | España |
| Institución: | Universidad de Barcelona |
| Repositorio: | Dipòsit Digital de la UB |
| OAI Identifier: | oai:diposit.ub.edu:2445/131908 |
| Acceso en línea: | https://hdl.handle.net/2445/131908 http://hdl.handle.net/10803/666611 |
| Access Level: | acceso abierto |
| Palabra clave: | Metazous Proteïnes Anàlisi de proteïnes Integrines Metazoa Proteins Analysis of proteins Integrins |
| Sumario: | [eng] Animal multicellular structure depends on three general mechanisms: cell adhesion, cell communication and cell differentiation. Some of these mechanisms were thought to be specific to animals. Genome sequencing of unicellular organisms that are closely related to animals has revealed that the unicellular ancestor of all animals already contained homologs of some of the genes that have a role in cell adhesion, cell communication and cell differentiation. Studying the function of these genes in their unicellular context is the way to infer the function they could have had in the ancestor, thus, helping us to understand how animals originated. Capsaspora owczarzaki, a filopodiated amoeba, is the closer unicellular relative of animals that contains key elements of the major cell-matrix adhesion system that animal cells use: the integrin adhesome. Thus, Capsaspora is an ideal model to study the origin of the integrin adhesome. However, the tools that would make feasible the experiments to study the function of these genes are not available. Therefore, this work has focused on developing molecular tools to work with Capsaspora, as well as understanding with other means the function of proteins related to the integrin adhesome. First, the life cycle of Capsaspora has been described at a cellular and molecular level. This revealed that the life stages of Capsaspora are transcriptomically regulated. Characterisation of its epigenome has revealed that these stages are regulated by some mechanisms which are shared with animals. In order to perform genetic studies, a transfection protocol based in calcium-phosphate precipitation has been established for Capsaspora. It was successfully used to label different structures of the cell with fluorescent tags, one of such allowed to visualise with unprecedented detail the filopodia in Capsaspora. As a first approach to the function of the integrin adhesome in Capsaspora, antibodies were designed to localise integrin adhesome proteins in the cell, and an adhesion assay was set up to analyse the adhesive potential of Capsaspora. Localisation of proteins in the filopodia and adhesion to fibronectin suggest that the integrin adhesome proteins in Capsaspora could be involved in adhesive function as in animals. The tools developed during this work are the first steps towards turning Capsaspora into a model organism to study the function of proteins involved in animal multicellularity. |
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