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Identification of multiprotein complexes controlling mitosis and cytokinesis in drosophila

Reference	BB/C508926/1
Principal Investigator / Supervisor	Professor David Glover
Co-Investigators / Co-Supervisors	Dr Pier Paolo D'Avino, Professor Ernest Laue, Professor Kathryn Lilley
Institution	University of Cambridge
Department	Genetics
Funding type	Research
Value (£)	498,557
Status	Completed
Type	Research Grant
Start date	01/11/2004
End date	31/01/2008
Duration	39 months

Abstract

The complex processes of mitosis and cytokinesis require the co-ordinated function of many multiprotein complexes. Over the past 20 years David Glover's laboratory has studied genes essential for mitosis and meiosis in Drosophila melanogaster using a combination of genetic and molecular approaches. This has led to the identification and analysis of several crucial components of the cell division machinery. In addition, a recent genome-wide screen in cultured Drosophila cells by Glover's laboratory in collaboration with Cyclacel Ltd has identified 400 novel genes that, when silenced by RNAi, lead to a change in mitiotic index. To characterise interactions between these mitotic regulators, it is proposed to use the TAP (Tandem Affinity Purification) methods recently developed in S. cerevisiae and successfully used for systematic genome-wide characterisation of native protein complexes. Proteins will be fused to two diverse affinity tags and introduced into Drosophila cells. The isolation of the tagged protein with its partners will be accomplished through two gentle affinity purification steps and components of the complex identified by mass spectrometry. We will first develop and optimise TAP tools and protocols for purifying known, partially characterised mitiotic complexes and then tag up to 400 cell cycle regulatory proteins identified in our functional screen in order to initiate complex characterisation. The protein tag will also enable the sub-cellular localisation of the complex to be determined. The resulting knowledge of function, localisation and protein partners for any particular cell cycle regulatory protein will allow the building of a functional network that will offer insight into the workings of the molecular machinery of cell division in metazoans.

Summary

unavailable

Committee	Closed Committee - Genes & Developmental Biology (GDB)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	Proteomics and Cell Function (PCF) [2003-2004]
Funding Scheme	X – not Funded via a specific Funding Scheme

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