Force generation at microtubule ends: An in vitro approach to cortical interactions. Liedewij Laan
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1 Force generation at microtubule ends: An in vitro approach to cortical interactions Liedewij Laan
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3 Force generation at microtubule ends: An in vitro approach to cortical interactions PROEFSCHRIFT TER VERKRIJGING VAN DE GRAAD VAN DOCTOR AAN DE UNIVERSITEIT LEIDEN, OP GEZAG VAN DE RECTOR MAGNIFICUS PROF. MR. P.F. VAN DER HEIJDEN, VOLGENS BESLUIT VAN HET COLLEGE VOOR PROMOTIES TE VERDEDIGEN OP WOENSDAG 10 JUNI 2009 TE KLOKKE UUR DOOR LIEDEWIJ LAAN GEBOREN TE HENGELO (O) IN 1980
4 Promotiecommissie: Promotor: Overige leden: Prof. dr. M. Dogterom Prof. dr. R. D. Vale (University of California San Francisco, USA) Dr. S. Grill (MPI-CBG/MPI-PKS, Dresden, Germany) Prof. dr. R. H. Medema (Universiteit Utrecht) Prof. dr. C. Dekker (Technische Universiteit Delft) Prof. dr. H. Schiessel Prof. dr. T. Schmidt Prof. dr. J. M. van Ruitenbeek Force generation at microtubule ends: An in vitro approach to cortical interactions 2009 by Liedewij Laan. All rights reserved. Cover: Laura van Uitert The work described in this thesis was performed at the FOM Institute for Atomic and Molecular Physics (AMOLF), Science Park 113, 1098 XG Amsterdam. This work is part of the research programme of the Stichting voor Fundamenteel Onderzoek der Materie (FOM), which is financially supported by the Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO). It is also partly funded by the Human Frontiers Science Program. ISBN: A digital version of this thesis can be downloaded from Printed copies can be obtained by addressing the library at the FOM institute for Atomic and Molecular Physics (AMOLF): library@amolf.nl, Science Park 114, 1098 XG, Amsterdam, the Netherlands. Printed in the Netherlands by Ponsen & Looijen BV graphical company, Wageningen. 4
5 What I cannot create, I do not understand, Richard Feynman
6 This thesis is partly based on the following articles: Marileen Dogterom, Julien Husson, Liedewij Laan, E. Laura Munteanu, Christian Tischer Microtubule forces and organization, (Chapter 1, 3, 5) Cell motility / ed. P. Lenz. - New York : Springer, - pp (Springer series in Biological and Medical Physics, Biomedical Engineering) (2007) Liedewij Laan and Marileen Dogterom In vitro assays to study force generation at dynamic microtubule ends, (Chapter 2) (in preparation: invited contribution to Methods in Cell Biology) Julien Husson, Liedewij Laan, Marileen Dogterom Force-generation by microtubule bundles, (Chapter 3) Biophysical Reviews and Letters, in press (2009) Liedewij Laan*, Julien Husson*, E. Laura Munteanu, Jacob W. J. Kerssemakers, Marileen Dogterom Force generation and dynamic instability of microtubule bundles, (Chapter 3) Proc Natl Acad Sci USA, 105: (2008) (*equal contribution) Liedewij Laan, Julien Husson, Martijn van Duijn, Ronald D. Vale, Samara L. Reck-Peterson, Marileen Dogterom Cortex -attached dynein regulates and pulls on shrinking microtubule ends in vitro, (Chapter 4) (under review) Liedewij Laan, Nenad Pavin, Guillaume Romet-Lemonne, Frank Jülicher, Marileen Dogterom Reliable centering of dynamic MT asters in microfabricated chambers by pulling forces, (Chapter 5) (in preparation) Peter Bieling*, Liedewij Laan*, Henry Schek, E. Laura Munteanu, Linda Sandblad, Marileen Dogterom, Damian Brunner, Thomas Surrey Reconstitution of a microtubule plus-end tracking system in vitro, (Chapter 6) Nature 450, (2007) (*equal contribution) Other papers by this author: E. Laura Munteanu, Liedewij Laan, Damian Brunner, Thomas Surrey, Marileen Dogterom Regulation of microtubule dynamics, in vitro, by the autonomous microtubule-end tracker Mal3, (to be submitted) Liedewij Laan, Laura Munteanu, Jacob Kerssemakers, Marileen Dogterom Meten aan microbuisjes met moleculaire resolutie, Nederlands Tijdschrift voor Natuurkunde (2006) (Dutch physics journal) Jacob W. J. Kerssemakers, E. Laura Munteanu, Liedewij Laan, Tim Noetzel, Marcel Janson and Marileen Dogterom Assembly dynamics of microtubules at molecular resolution, Nature 442, (2006)
7 Contents Introduction Internal organization of the cell Microtubules in cellular organization Force generation in cellular organization Thesis outline...27 Mimicking microtubule interactions at the cell boundary with functionalized microfabricated structures Microfabrication Activation of gold barriers Assays with microfabricated structures Acknowledgements...45 Force generation and dynamic instability of microtubule bundles Introduction Measurement of bundle forces and dynamics Simulations on microtubule bundle dynamics Discussion Materials and Methods Acknowledgements...62 Cortical dynein regulates and pulls on dynamic microtubule ends in vitro Introduction Fluorescence microscopy experiments Optical trap experiments Discussion Materials and Methods Acknowledgements
8 Reliable centering of dynamic microtubule asters in microfabricated chambers by pulling forces Introduction Experimental results Theory Comparison of experiments and theory Discussion Materials and methods Acknowledgements Reconstitution of a microtubule plus-end tracking system in vitro Introduction Results on end-tracking in vitro Discussion Materials and Methods Acknowledgements Concluding remarks and additional research directions Positioning processes based on microtubule pulling forces Regulation of microtubule dynamics by a combination of force and +TIPs Microtubule capture by non-motor proteins Towards the role of MT-based transport in cortical pattern formation Acknowledgements Bibliography Summary Samenvatting Dankwoord Curriculum Vitae
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