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In vitro reconstitution of the S. pombe interphase microtubule cytoskeleton

Objective

The research proposed here intends to explore processes governed by microtubule (MT) plus end accumulating proteins (+TIPs) and interdependencies between these proteins for their localisation. The well defined but relatively simple interphase MT cytoskeleton of Schitzosaccharomyces pombe (S. pombe) has proven to be an excellent model for MT plus end studies. In interphase, MTs are involved in defining sites of polarised growth as well as nuclear positioning by generating pushing forces against the cell cortex. The mechanochemical principles governing this process as well as the likely interplay between +TIPs are as yet incompletely understood. This proposal focuses on elucidating the interdependencies between the various evolutionary conserved +TIPs using (purified proteins of) S. pombe in both in vivo and in vitro experiments. Initial experiments will determine the function of the recently characterised class of length-dependent MT-depolymerases (kinesin-8s) in MT plus end behaviour in S. pombe. Combined with already characterised +TIPs such as Mal3, Tip1, and Tea2, a complete set of proteins, governing all observed MT dynamic behaviours seen in vivo, may now be available. These will be used in an attempt to recreate the first fully functional MT plus end in vitro. Expanding on a novel microscopy set-up created by the Surrey laboratory (EMBL, Heidelberg, Germany, unpublished) we hope to determine the interplay between +TIPs for MT plus end localisation. In addition, we want to investigate the individual as well as the combined influence of these +TIPs on MT dynamic behaviour in vitro. Skills acquired during this research at the interphase of cell biology and biophysics using both in vitro and in vivo techniques will be highly advantageous in obtaining an independent position after the fellowship in these developing fields of biological research.

Field of science

  • /natural sciences/biological sciences/cell biology
  • /natural sciences/biological sciences/biophysics
  • /natural sciences/biological sciences/biochemistry/biomolecules/proteins

Call for proposal

FP7-PEOPLE-2007-2-1-IEF
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Funding Scheme

MC-IEF - Intra-European Fellowships (IEF)

Coordinator

EUROPEAN MOLECULAR BIOLOGY LABORATORY
Address
Meyerhofstrasse 1
69117 Heidelberg
Germany
Activity type
Research Organisations
EU contribution
€ 158 694,85
Administrative Contact
Damian Brunner (Dr.)