How does the autism-related protein Shank contribute to the regulation of neuronal spine plasticity?
Lead Research Organisation:
University of Liverpool
Department Name: Institute of Integrative Biology
Abstract
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Organisations
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| BB/M011186/1 | 30/09/2015 | 31/03/2024 | |||
| 1945241 | Studentship | BB/M011186/1 | 30/09/2017 | 30/03/2022 | Emily Wang |
| NE/W503083/1 | 31/03/2021 | 30/03/2022 | |||
| 1945241 | Studentship | NE/W503083/1 | 30/09/2017 | 30/03/2022 | Emily Wang |
| Description | The work has so far focused on further understanding the structure of the N-terminal region of the Shank3 protein to help further understanding of how this protein has a regulatory function in dendritic spines. The N-terminal region of Shank3 is composed of two domains, the Shank/Prosap N-terminal domain (SPN) and Ankyrin Repeat (ANK) domains that are associated through a polar interface. We have found that some significant autism-associated mutations that are found in this N-terminal region cause significant de-stabalisation of the N-terminal domains. Therefore we have hypothesised that the stability of the N-terminal region is critical for Shank3 neuronal function. NMR analysis of the very N-terminal domain, the SPN domain, has shown that it is intrinsically unstable when it is not in contact with the ANK domain, particularly in salt concentrations below 0.5M. NMR analysis, using mutations designed to disrupt the SPN-ANK interface , has shown that the separation of the SPN domain leads to its destabilisation. Moreover, single point mutations have dramatic effects on the N-terminal region, such as the autism-associated P141A, which causes destabilisation of the interface and reduces overall stability of the N-terminal region. These findings were confirmed through melting temperature data measured by CD and nanoDSF. From these findings, we propose an 'open-closed' model of the N-terminal region, where the SPN domain dynamically dissociates from the ANK domain, simultaneously leading to self-association and activation of the Shank3 binding sites. These two effects need to be taken into consideration when analysing autism-related mutations of Shank3. In addition, the previously solved interaction between the Shank3 N-terminal region and the Ras-family GTPases has been explored further and in association with our collaborators we have shown another interaction between the Shank3 N-terminal and the active K-Ras GTPase. |
| Exploitation Route | These findings contribute to the overall understanding of the Shank3 protein and this knowledge can be used to further determine regulatory mechanisms disrupted in the dendritic spines of neurons, that could contribute to disorders such as autism spectrum disorders. The structural information learnt can be aligned with observations seen in cell culture with mutations of Shank3 to fully understand how these mutations may be working in cells. In addition, this detailed structural knowledge of the domains of Shank3 could help later on with the development of potential drugs to help with neuronal disorders. |
| Sectors | Healthcare Pharmaceuticals and Medical Biotechnology |