Image: Drosophila female reproductive machinery, developing eggs (magenta), oviduct (cyan), nuclei (blue).

Global content and functional analysis of in vivo macromolecular complexes

Supervisor: Professor Tim Weil

Over the past decade, biomolecular condensates have emerged as a key concept in cellular organisation. These membraneless assemblies provide dynamic subcellular environments and often function as reaction crucibles. Condensates likely predated membranous organelles as the primordial organising principle which would have enabled early RNA based lifeforms to compartmentalise chemical processes. Despite extensive in vitro and in cellulo research, little is known about the composition of these fundamental structures and how they function in complex organisms. Condensates are also implicated in human diseases, especially those relating to aberrant protein aggregation, such as Alzheimer’s and Parkinson’s, or translational regulation, such as cancer.

This project aims to: (1) identify the complete mRNA content of in vivo condensates, (2) identify the complete protein content of in vivo condensates, and (3) establish the regulatory principles governing association with and disassociation from in vivo condensates.

To achieve these aims, novel flow cytometry will be used to separate biomolecular condensates from other cellular elements. This recently established, high throughput approach has the ability to gate condensates by size and acquires fluorescent images of particles simultaneously while sorting. Preliminary data from Drosophila oocytes and embryos shows that RNA sequencing and proteomics can be performed on sorted samples. Taking advantage of the toolkit available to Drosophila, this provides an unprecedented opportunity to analyse in vivo condensates experiencing different genetic, chemical, and environmental conditions. Together, this project will generate the most complete understanding of an in vivo macromolecular complex to date and will likely inform new therapeutic delivery methods for mRNA.

For more information, please visit Weil Lab or contact Prof Tim Weil.

References

Wilby EL and Weil TT, Relating the Biogenesis and Function of P Bodies in Drosophila to Human Disease. Genes. 2023; 14(9):1675. doi.org/10.3390/genes14091675

Roden C and Gladfelter AS RNA contributions to the form and function of biomolecular condensates. Nat Rev Mol Cell Biol. 2021 Mar;22(3):183-195. doi: 10.1038/s41580-020-0264-6.