Rotation 1: Using engineered protein systems for studying LLPS in the cellular environment
Lead Research Organisation:
University of Cambridge
Department Name: School of Biological Sciences
Abstract
BBSRC strategic theme: Understanding the rules of life
Liquid-liquid phase separation (LLPS) is a modern principle that involves compartmentalisation of proteins and nucleic acids into membraneless subcellular compartments, also termed as biomolecular condensates (BCs). Due to the integral role of LLPS in spatial and temporal regulation of biological processes, the role of BCs has been established in physiological contexts such as transcription regulation, protein degradation and neuronal synaptic signalling. However, recent studies have shown that corruption of this reversible natural process results in defective protein aggregation in pathological contexts such as neurodegenerative diseases. The aims of this project will therefore be to use designer LLPS-CTPRs to:
- characterise the physicochemical properties of BCs formed in cellulo
- understand the molecular mechanisms by which neurodegenerative proteins dysregulate the LLPS environment leading to irreversible aggregation.
In this project, we intend to utilize HeLa cell models transiently transfected with LLPS-CTPR proteins. Using these cell models, we will explore the properties of these condensates using live cell imaging via confocal fluorescence microscopy to monitor droplet structure and fusion; and FRAP, to characterize the dynamics of the proteins in the droplets. We will investigate novel LLPS-CTPRs to recruit disease-related proteins, such as alpha-synuclein and TDP-43, to try and consolidate how LLPS impacts their aberrant behaviour. Ultimately, we hope to determine if disease-related BCs can be targeted to the autophagy pathway. As aberrantly misfolded proteins are a pathological hallmark of neurodegenerative diseases, removal of these species through facilitated protein degradation may prove beneficial for therapeutic intervention.
Liquid-liquid phase separation (LLPS) is a modern principle that involves compartmentalisation of proteins and nucleic acids into membraneless subcellular compartments, also termed as biomolecular condensates (BCs). Due to the integral role of LLPS in spatial and temporal regulation of biological processes, the role of BCs has been established in physiological contexts such as transcription regulation, protein degradation and neuronal synaptic signalling. However, recent studies have shown that corruption of this reversible natural process results in defective protein aggregation in pathological contexts such as neurodegenerative diseases. The aims of this project will therefore be to use designer LLPS-CTPRs to:
- characterise the physicochemical properties of BCs formed in cellulo
- understand the molecular mechanisms by which neurodegenerative proteins dysregulate the LLPS environment leading to irreversible aggregation.
In this project, we intend to utilize HeLa cell models transiently transfected with LLPS-CTPR proteins. Using these cell models, we will explore the properties of these condensates using live cell imaging via confocal fluorescence microscopy to monitor droplet structure and fusion; and FRAP, to characterize the dynamics of the proteins in the droplets. We will investigate novel LLPS-CTPRs to recruit disease-related proteins, such as alpha-synuclein and TDP-43, to try and consolidate how LLPS impacts their aberrant behaviour. Ultimately, we hope to determine if disease-related BCs can be targeted to the autophagy pathway. As aberrantly misfolded proteins are a pathological hallmark of neurodegenerative diseases, removal of these species through facilitated protein degradation may prove beneficial for therapeutic intervention.
Organisations
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| BB/X010899/1 | 01/10/2023 | 30/09/2028 | |||
| 2888268 | Studentship | BB/X010899/1 | 01/10/2023 | 30/09/2027 | Belinda Agbetiameh |