Investigating the interaction between PHAF1 and the ATG8-family proteins in autophagy
Key information
Research topics
This is a summer student position supervised by Philippa Rosewell and Alina Ueffing in Sharon Tooze's lab.
Introduction to the science
Our lab is interested in understanding the molecular mechanisms that control autophagy, a cellular degradation pathway. Autophagy is evolutionarily conserved and its dysfunction is associated with a broad range of human pathologies, including neurodegenerative diseases such as Parkinson's Disease.
About the project
The evolutionary conserved protein PHAF1 has been shown to control life span and health span in C. elegans and to be involved in macroautophagy through interaction with BCAS and WIPI2. Interestingly, PHAF1 possesses several putative LC3-interacting regions (LIR), of which the most N-terminal one has been shown to be required for co-immunoprecipitation of LC3B with overexpressed PHAF1. Additionally, LC3B puncta formation could not be rescued with overexpression of PHAF1 lacking this LIR in C2C12 PHAF1 knockout cells, whereas WIPI2 puncta formation was restored [1]. However, whether this phenotype occurs in other cell lines (e.g. Hek293A) and whether PHAF1 interacts only with LC3B or also its paralogs remains to be elucidated. This project aims to investigate this interaction using both in-vitro approaches and/or cell biology and biochemistry. Overexpression of PHAF1 wildtype and LIR deficient versions will be applied to investigate interaction/colocalization with LC3B and its paralogs by confocal microscopy a well as co-immunoprecipitation. Additionally, affinities between different PHAF1 LIR peptides and LC3B and its paralogs will be determined to evaluate which paralog is defining for this interaction in cells.
Candidate background
The post holder should embody and demonstrate the Crick ethos and ways of working: bold, open and collegial. The candidate must be registered at a UK Higher Education Institution, studying in the UK and must have completed a minimum of two years’ undergraduate study in a relevant discipline, and on track to receive a final degree grade of 2:1 or 1. In addition, they should be able demonstrate the following experience and key competencies:
- This project would suit someone with studying molecular or cell biology or biochemistry - specifically with an interest in fundamental mechanisms of biological processes.
- Good knowledge in relevant scientific area(s)
- Good written and spoken communication skills
- Ability to work independently and also capable of interacting within a group
References
1. Franco-Romero, A., Morbidoni, V., Milan, G., Sartori, R., Wulff, J., Romanello, V., . . . Sandri, M. (2024)
C16ORF70 /MYTHO promotes healthy aging in C.elegans and prevents cellular senescence in mammals.
Journal of Clinical Investigation 134: e165814. PubMed abstract
2. Leduc-Gaudet, J.P., Franco-Romero, A., Cefis, M., Moamer, A., Broering, F.E., Milan, G., . . . Gouspillou, G. (2023)
MYTHO is a novel regulator of skeletal muscle autophagy and integrity.
Nature Communications 14: 1199. PubMed abstract