what principles govern the formation of the mitotic spindle? how do they interact to ensure...
TRANSCRIPT
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