Force generation at microtubule ends : An in vitro approach to cortical interactions.
Laan, L.
Citation
Laan, L. (2009, June 10). Force generation at microtubule ends : An in vitro approach to
Force generation at microtubule ends:
An in vitro approach to cortical interactions
Liedewij Laan
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 15.00 UUR
DOOR
L
IEDEWIJL
AANGEBOREN TE HENGELO (O) IN 1980
Promotiecommissie:
Promotor: Prof. dr. M. Dogterom
Overige leden: 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: 978-90-77209-33-2
A digital version of this thesis can be downloaded from http://ub.leidenuniv.nl. Printed copies can be
What I cannot create, I do not understand, Richard Feynman
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. 93-115
(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, 105:8920-8925 (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, 1100-1105 (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
7
Contents
Introduction ...9
1.1 Internal organization of the cell ...10
1.2 Microtubules in cellular organization...12
1.3 Force generation in cellular organization ...18
1.4 Thesis outline ...27
Mimicking microtubule interactions at the cell boundary with functionalized microfabricated structures ...29
2.1 Microfabrication...30
2.2 Activation of gold barriers ...37
2.3 Assays with microfabricated structures...39
2.4 Acknowledgements...45
Force generation and dynamic instability of microtubule bundles ...47
3.1 Introduction ...48
3.2 Measurement of bundle forces and dynamics ...49
3.3 Simulations on microtubule bundle dynamics...55
3.4 Discussion...57
3.5 Materials and Methods...60
3.6 Acknowledgements...62
‘Cortical’ dynein regulates and pulls on dynamic microtubule ends in vitro...63
4.1 Introduction ...64
4.2 Fluorescence microscopy experiments...64
4.3 Optical trap experiments ...70
4.4 Discussion...75
4.5 Materials and Methods...79
4.6 Acknowledgements. ...84
Reliable centering of dynamic microtubule asters in microfabricated chambers
by pulling forces ...85
5.1 Introduction ...86
5.2 Experimental results ...87
5.3 Theory ...91
5.4 Comparison of experiments and theory...97
5.5 Discussion...98
5.6 Materials and methods ...100
5.7 Acknowledgements...103
Reconstitution of a microtubule plus-end tracking system in vitro ...105
6.1 Introduction ...106
6.2 Results on end-tracking in vitro ...106
6.3 Discussion...117
6.4 Materials and Methods...120
6.5 Acknowledgements...128
Concluding remarks and additional research directions ...129
7.1 Positioning processes based on microtubule pulling forces ...130
7.2 Regulation of microtubule dynamics by a combination of force and +TIPs ....131
7.3 Microtubule capture by non-motor proteins ...134
7.4 Towards the role of MT-based transport in cortical pattern formation...139
7.5 Acknowledgements...146
Bibliography ...147
Summary ...163
Samenvatting ...167
