Investigation into how Cdc42 is activated to define the apical domain of epithelial cells in Drosophila
Lead Research Organisation:
University of Cambridge
Department Name: Gurdon Institute
Abstract
Theme: World-Class Underpinning Bioscience
The small GTPase Cdc42 is a highly conserved key regulator of apicobasal polarity. It plays an important role in localising apical determinants, such as Par6 and aPKC, and tight control of its activity is crucial for establishment and maintenance of epithelial polarity. Activity of Cdc42 is controlled by guanine nucleotide exchange factors (GEFs) and GTPase activating proteins (GAPs) that promote activation or inactivation, respectively. Our lab has identified putative Cdc42 GEFs in Drosophila, including a GEF that phenocopies Cdc42 mutants in the follicular epithelium. In this project I would like to examine the activity of this GEF, CG42674, on Cdc42 and other members of the Rho GTPase family. In addition, I want to determine if the GEF is localised to the apical domain in follicle cells, where active Cdc42 (Cdc42-GTP) is found. I also want to take an unbiased approach to elucidate how CG42674 is localised. It would be interesting to identify the protein responsible for localising the GEF as it is upstream of Cdc42, a master regulator of polarity. To study this question, I will make use of novel biotin-based proximity labelling technique TurboID. This will be the first time that TurboID is used to answer a biological question in Drosophila. I would also like to examine the role that other putative Cdc42 GEFs play in polarity maintenance of various epithelia. Using TurboID I could identify potential interacting proteins for these other GEFs in their relevant tissues and discover how they are localised to the correct domain where they can activate Cdc42. I aim to find common mechanisms across various epithelia in how the apical domain is maintained through localised activity of the relevant dc42 GEF.
The small GTPase Cdc42 is a highly conserved key regulator of apicobasal polarity. It plays an important role in localising apical determinants, such as Par6 and aPKC, and tight control of its activity is crucial for establishment and maintenance of epithelial polarity. Activity of Cdc42 is controlled by guanine nucleotide exchange factors (GEFs) and GTPase activating proteins (GAPs) that promote activation or inactivation, respectively. Our lab has identified putative Cdc42 GEFs in Drosophila, including a GEF that phenocopies Cdc42 mutants in the follicular epithelium. In this project I would like to examine the activity of this GEF, CG42674, on Cdc42 and other members of the Rho GTPase family. In addition, I want to determine if the GEF is localised to the apical domain in follicle cells, where active Cdc42 (Cdc42-GTP) is found. I also want to take an unbiased approach to elucidate how CG42674 is localised. It would be interesting to identify the protein responsible for localising the GEF as it is upstream of Cdc42, a master regulator of polarity. To study this question, I will make use of novel biotin-based proximity labelling technique TurboID. This will be the first time that TurboID is used to answer a biological question in Drosophila. I would also like to examine the role that other putative Cdc42 GEFs play in polarity maintenance of various epithelia. Using TurboID I could identify potential interacting proteins for these other GEFs in their relevant tissues and discover how they are localised to the correct domain where they can activate Cdc42. I aim to find common mechanisms across various epithelia in how the apical domain is maintained through localised activity of the relevant dc42 GEF.
Organisations
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| BB/M011194/1 | 01/10/2015 | 31/03/2024 | |||
| 1943864 | Studentship | BB/M011194/1 | 01/10/2017 | 30/06/2020 | Erinn Los |
| Description | Honorary Benefactor's Scholarship |
| Amount | £1,300 (GBP) |
| Organisation | University of Cambridge |
| Department | St John's College |
| Sector | Academic/University |
| Country | United Kingdom |
| Start | 10/2017 |
| End | 09/2020 |
| Description | Open day at the Gurdon Insitute |
| Form Of Engagement Activity | Participation in an open day or visit at my research institution |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Public/other audiences |
| Results and Impact | Open day at our research institute, the Gurdon Institute, led to engagement with the general public on the research that is being carried out in the institute. I've discussed both my own work and that of others in the building, as well as the importance of scientific research. |
| Year(s) Of Engagement Activity | 2018,2019 |
| Description | Participation in Scientists' Collaborative Project with Educators (SCoPE) |
| Form Of Engagement Activity | Engagement focused website, blog or social media channel |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Schools |
| Results and Impact | Teachers and Gurdon Institute scientists co-created four innovative teaching 'toolkits' to use in biology classrooms across the UK. These toolkits are part of our public engagement programme and they are free to use by teachers in classrooms around the United Kingdom. The project I worked on resulted in a team-based puzzle-solving game for reviewing transcription and translation and teaching the students about gene editing. |
| Year(s) Of Engagement Activity | 2019,2020 |
| URL | https://scopegurdoninstitute.co.uk/unlockinggeneediting |