Standard

The dynamics of microtubule stability : A reconstitution of regulated microtubule assembly under force. / Kok, Maurits.

2019-10 ed. Delft-Leiden : Casimir PhD Series, 2019. 167 p.

Research output: ThesisDissertation (TU Delft)Scientific

Harvard

APA

Vancouver

Author

BibTeX

@phdthesis{5cb3dd6078ab4272a8207a754c2cc6a0,
title = "The dynamics of microtubule stability: A reconstitution of regulated microtubule assembly under force",
abstract = "The microtubule cytoskeleton is an intracellular polymer network involved in cell shape, cell motility, and cell division. Microtubules are self-assembling, dynamic polymers composed of tubulin proteins that alternate between phases of growth and shrinkage, a behaviour known as “dynamic instability”. A key feature of microtubules is their ability to generate pushing and pulling forces through controlled assembly and disassembly, guiding for example cell division. The stability of microtubules is largely governed by the progressive hydrolysis of incorporated tubulin dimers. In this thesis we sought to resolve microtubule assembly under force in the presence of regulatory proteins. Through the in vitro reconstitution of microtubule growth against novel micro-fabricated barriers, we studied the biochemical changes leading up to microtubule destabilization. We found that the resulting microtubule lifetimes can be understood with a simple phenomenological 1D model based on noisy microtubule growth and a single hydrolysis rate. Additionally, we explored the implementation of light-inducible protein-protein interactions to gain spatiotemporal control over microtubule function in particular and over in vitro reconstitutions in general. As a proof-of-principle, we developed a light-inducible microtubule gliding assay based on the reversible recruitment of motor proteins to a surface.",
keywords = "Microtubules, Optogenetics, Micro-fabrication, Reconstitution, EB3, CLASP, Monte Carlo simulations, Motor proteins",
author = "Maurits Kok",
year = "2019",
month = "4",
day = "24",
doi = "10.4233/uuid:5cb3dd60-78ab-4272-a820-7a754c2cc6a0",
language = "English",
isbn = "978-90-8593-392-2",
series = "Casimir PhD Series",
publisher = "Casimir PhD Series",
number = "2019-10",
edition = "2019-10",
school = "Delft University of Technology",

}

RIS

TY - THES

T1 - The dynamics of microtubule stability

T2 - A reconstitution of regulated microtubule assembly under force

AU - Kok, Maurits

PY - 2019/4/24

Y1 - 2019/4/24

N2 - The microtubule cytoskeleton is an intracellular polymer network involved in cell shape, cell motility, and cell division. Microtubules are self-assembling, dynamic polymers composed of tubulin proteins that alternate between phases of growth and shrinkage, a behaviour known as “dynamic instability”. A key feature of microtubules is their ability to generate pushing and pulling forces through controlled assembly and disassembly, guiding for example cell division. The stability of microtubules is largely governed by the progressive hydrolysis of incorporated tubulin dimers. In this thesis we sought to resolve microtubule assembly under force in the presence of regulatory proteins. Through the in vitro reconstitution of microtubule growth against novel micro-fabricated barriers, we studied the biochemical changes leading up to microtubule destabilization. We found that the resulting microtubule lifetimes can be understood with a simple phenomenological 1D model based on noisy microtubule growth and a single hydrolysis rate. Additionally, we explored the implementation of light-inducible protein-protein interactions to gain spatiotemporal control over microtubule function in particular and over in vitro reconstitutions in general. As a proof-of-principle, we developed a light-inducible microtubule gliding assay based on the reversible recruitment of motor proteins to a surface.

AB - The microtubule cytoskeleton is an intracellular polymer network involved in cell shape, cell motility, and cell division. Microtubules are self-assembling, dynamic polymers composed of tubulin proteins that alternate between phases of growth and shrinkage, a behaviour known as “dynamic instability”. A key feature of microtubules is their ability to generate pushing and pulling forces through controlled assembly and disassembly, guiding for example cell division. The stability of microtubules is largely governed by the progressive hydrolysis of incorporated tubulin dimers. In this thesis we sought to resolve microtubule assembly under force in the presence of regulatory proteins. Through the in vitro reconstitution of microtubule growth against novel micro-fabricated barriers, we studied the biochemical changes leading up to microtubule destabilization. We found that the resulting microtubule lifetimes can be understood with a simple phenomenological 1D model based on noisy microtubule growth and a single hydrolysis rate. Additionally, we explored the implementation of light-inducible protein-protein interactions to gain spatiotemporal control over microtubule function in particular and over in vitro reconstitutions in general. As a proof-of-principle, we developed a light-inducible microtubule gliding assay based on the reversible recruitment of motor proteins to a surface.

KW - Microtubules

KW - Optogenetics

KW - Micro-fabrication

KW - Reconstitution

KW - EB3

KW - CLASP

KW - Monte Carlo simulations

KW - Motor proteins

U2 - 10.4233/uuid:5cb3dd60-78ab-4272-a820-7a754c2cc6a0

DO - 10.4233/uuid:5cb3dd60-78ab-4272-a820-7a754c2cc6a0

M3 - Dissertation (TU Delft)

SN - 978-90-8593-392-2

T3 - Casimir PhD Series

PB - Casimir PhD Series

CY - Delft-Leiden

ER -

ID: 52298977