1. What principles govern the formation of the mitotic spindle? How do they interact to ensure robustness and flexibility to the system? Why is the spindle the shape it is? Cell division – The Wakefield Lab

2. qualitative synthetic quantitative analytical System perturbation (genetics et al.) Proteomics Software engineering Biochemistry Bioinformatics What is our approach? Mathematical modelling early embryos Observation (cell biology) 13 rounds of mitosis within 3 hrs Maternally produced protein Syncytial and therefore synchronous Can collect grammes within a week

3. What proteins that bind MTs are important for cell division? biochemically isolate Drosophila embryonic MAPs Identify via MS and categorise controlmsd1 RNAi msd3 RNAi live observation Correlation categorisation Characterise novel proteins Hughes et al., (2008) PLoS Biol

4. Applying MS data to protein interaction networks Use MS data to produce a list of interspecies MAPs Generate extended network of putative MAPs Rank using PCA/Random forest likelihood statistics Generate interesting hypotheses characterise candidates

5. What proteins are part of mitotic MAP complexes in vivo, and what PTMs are important for their function? cytokinesis kinetochore:k-MT attachment AURORA B SURVIVIN BOREALIN INCENP chromosome condensation spindle organisation metaphase anaphase 49 171 63 7 65 13 Metaphase Anaphase Mass spec and analyse characterise candidates Purify via GFP-TRAP Identify phosphorylation and SUMOylation sites

6. Acknowledgements Wakefield Lab (Past) Julian Hughes Katherine Fisher (Present) Faisal Khan Pete Jones Dan Hayward Jack Chen Simon Li Collaborations  Mass Spectrometry Nicole Zitzmann (Oxford) Hannah Florance (Exeter)  Bioinformatics Charlotte Deane (Oxford)

7. What principles govern the formation of the mitotic spindle? How do they interact to ensure robustness and flexibility to the system? Why is the spindle the shape it is? all principles / processes