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

qualitativesynthetic

quantitativeanalytical

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

What proteins that bind MTs are important for cell division?

biochemically isolate Drosophila embryonic MAPs

Identify via MS and categorisecontrol msd1 RNAi msd3 RNAi

live observationCorrelationcategorisation

Characterise novel proteins

Hughes et al., (2008) PLoS Biol

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 statisticsGenerate

interesting hypothesescharacterise

candidates

What proteins are part of mitotic MAP complexes in vivo, and what PTMs are important for their function?

cytokinesis

kinetochore:k-MTattachment

AURORA B

SU

RV

IVINBOREALIN

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

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)

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