Structural Analysis of Centrosome Biogenesis

Most eukaryotic cells are born with a single centrosome that plays an important part in many aspects of cellular organization. Centrosomes are an excellent model for studying organelle biogenesis because, just like the DNA, they duplicate precisely once during each cell cycle. In the early Drosophila embryo, hundreds of centrosomes synchronously assemble every few minutes as the embryos rapidly progress through repeated cycles of division. 

Although centrosomes are complex nanomachines comprising >400 proteins, only ~10 proteins are absolutely essential for centrosome biogenesis. Thus, to understand the principles that allow these embryos to coordinately assemble so many centrosomes at the right time, in the right place, and then grow them to the right size, we need to understand how these proteins interact with each other, and how these interactions are regulated. This project involves using protein prediction software (e.g. AlphaFold2, Rosetta) to identify putative interactions and then using various approaches (biochemistry, CryoEM, in vitro reconstitution) to validate them (NIH). Once validated, the functional significance of these interactions will be tested using live-cell imaging in the early Drosophila embryo (Oxford). See publications below for two examples of previous collaborations between the NIH and Oxford groups.

Feng et al., Structural basis for mitotic centrosome assembly in flies. Cell, 2017.

Conduit et al., The centrosome-specific phosphorylation of Cnn by Polo/PLK1 drives Cnn scaffold assembly and centrosome maturation. Dev. Cell., 2014.