document

Progress in Engineering Synthetic Cells and Cell-Free Systems

Dogterom, A.M. (author), Kamat, Neha P. (author), Jewett, Michael C. (author), Adamala, Katarzyna P. (author)

contribution to periodical 2024

document

Reconstituting actin-microtubule crosstalk

Dogterom, A.M. (author)

journal article 2023

document

High-speed imaging of giant unilamellar vesicle production in cDICE

Van de Cauter, L. (author), Jawale, Y.K. (author), Tam, D.S.W. (author), Baldauf, L. (author), Koenderink, G.H. (author), Dogterom, A.M. (author), Ganzinger, Kristina A. (author)

journal article 2023

document

Stable kinetochore-microtubule attachment requires loop-dependent Ndc80-Ndc80 binding

Polley, Soumitra (author), Müschenborn, Helen (author), Terbeck, Melina (author), De Antoni, Anna (author), Vetter, Ingrid R. (author), Dogterom, A.M. (author), Musacchio, Andrea (author), Volkov, V. (author), Huis in 't Veld, Pim J. (author)

During cell division, kinetochores link chromosomes to spindle microtubules. The Ndc80 complex, a crucial microtubule binder, populates each kinetochore with dozens of copies. Whether adjacent Ndc80 complexes cooperate to promote microtubule binding remains unclear. Here we demonstrate that the Ndc80 loop, a short sequence that interrupts the...

journal article 2023

document

Estimation of microtubule-generated forces using a DNA origami nanospring

Nick Maleki, A. (author), Huis In 't Veld, Pim J. (author), Akhmanova, Anna (author), Dogterom, A.M. (author), Volkov, V. (author)

Microtubules are dynamic cytoskeletal filaments that can generate forces when polymerizing and depolymerizing. Proteins that follow growing or shortening microtubule ends and couple forces to cargo movement are important for a wide range of cellular processes. Quantifying these forces and the composition of protein complexes at dynamic...

journal article 2023

document

CSPP1 stabilizes growing microtubule ends and damaged lattices from the luminal side

van den Berg, Cyntha M. (author), Volkov, V. (author), Schnorrenberg, Sebastian (author), Huang, Ziqiang (author), Stecker, Kelly E. (author), Grigoriev, Ilya (author), Gilani, Sania (author), Frikstad, Kari Anne M. (author), Dogterom, A.M. (author)

Microtubules are dynamic cytoskeletal polymers, and their organization and stability are tightly regulated by numerous cellular factors. While regulatory proteins controlling the formation of interphase microtubule arrays and mitotic spindles have been extensively studied, the biochemical mechanisms responsible for generating stable...

journal article 2023

document

Present and future of synthetic cell development

Adamala, Katarzyna P. (author), Dogterom, A.M. (author), Elani, Yuval (author), Schwille, Petra (author), Takinoue, Masahiro (author), Tang, T. Y.Dora (author)

Scientists are captivated by the prospect of creating a fully synthetic cell, offering the potential to revolutionize biology, medicine and biotechnology. In this Viewpoint, a panel of experts discusses the definitions of a synthetic cell and highlights current achievements, challenges and future opportunities of building such systems.

journal article 2023

document

Strain stiffening of Ndc80 complexes attached to microtubule plus ends

Schwietert, Felix (author), Volkov, V. (author), Huis in ’t Veld, Pim J. (author), Dogterom, A.M. (author), Musacchio, Andrea (author), Kierfeld, Jan (author)

In the mitotic spindle, microtubules attach to chromosomes via kinetochores. The microtubule-binding Ndc80 complex is an integral part of kinetochores, and is essential for kinetochores to attach to microtubules and to transmit forces from dynamic microtubule ends to the chromosomes. The Ndc80 complex has a rod-like appearance with globular...

journal article 2022

document

Cross-linkers at growing microtubule ends generate forces that drive actin transport

Alkemade, C. (author), Wierenga, H. (author), Volkov, V. (author), López, Magdalena Preciado (author), Akhmanova, Anna (author), ten Wolde, Pieter Rein (author), Dogterom, A.M. (author), Koenderink, G.H. (author)

The actin and microtubule cytoskeletons form active networks in the cell that can contract and remodel, resulting in vital cellular processes such as cell division and motility. Motor proteins play an important role in generating the forces required for these processes, but more recently the concept of passive cross-linkers being able to...

journal article 2022

document

Multivalent interactions facilitate motor-dependent protein accumulation at growing microtubule plus-ends

Maan, R. (author), Reese, L. (author), Volkov, V. (author), King, M.R. (author), van der Sluis, E.O. (author), Andrea, N. (author), Evers, W.H. (author), Jakobi, A. (author), Dogterom, A.M. (author)

Growing microtubule ends organize end-tracking proteins into comets of mixed composition. Here using a reconstituted fission yeast system consisting of end-binding protein Mal3, kinesin Tea2 and cargo Tip1, we found that these proteins can be driven into liquid-phase droplets both in solution and at microtubule ends under crowding conditions....

journal article 2022

document

Shaping Liposomes by Cell-Free Expressed Bacterial Microtubules

Kattan, J.M. (author), Doerr, A. (author), Dogterom, A.M. (author), Danelon, C.J.A. (author)

Genetic control over a cytoskeletal network inside lipid vesicles offers a potential route to controlled shape changes and DNA segregation in synthetic cell biology. Bacterial microtubules (bMTs) are protein filaments found in bacteria of the genus Prosthecobacter. They are formed by the tubulins BtubA and BtubB, which polymerize in the...

journal article 2021

document

Building a community to engineer synthetic cells and organelles from the bottom-up

Staufer, Oskar (author), DE LORA, Jacqueline A. (author), Bailoni, Eleonora (author), Bazrafshan, Alisina (author), Benk, Amelie S. (author), Jahnke, Kevin (author), Manzer, Zachary A. (author), Dogterom, A.M. (author), Lopez, G.D.H. (author)

Employing concepts from physics, chemistry and bioengineering, 'learning-by-building' approaches are becoming increasingly popular in the life sciences, especially with researchers who are attempting to engineer cellular life from scratch. The SynCell2020/21 conference brought together researchers from different disciplines to highlight...

journal article 2021

document

Nonspherical Coacervate Shapes in an Enzyme-Driven Active System

Spoelstra, W.K. (author), van der Sluis, E.O. (author), Dogterom, A.M. (author), Reese, L. (author)

Coacervates are polymer-rich droplets that form through liquid-liquid phase separation in polymer solutions. Liquid-liquid phase separation and coacervation have recently been shown to play an important role in the organization of biological systems. Such systems are highly dynamic and under continuous influence of enzymatic and chemical...

journal article 2020

document

Spatiotemporal control of coacervate formation within liposomes

Deshpande, S.R. (author), Brandenburg, Frank (author), Lau, A.W.H.R. (author), Last, M.G.F. (author), Spoelstra, W.K. (author), Reese, L. (author), Wunnava, Sreekar (author), Dogterom, A.M. (author), Dekker, C. (author)

Liquid-liquid phase separation (LLPS), especially coacervation, plays a crucial role in cell biology, as it forms numerous membraneless organelles in cells. Coacervates play an indispensable role in regulating intracellular biochemistry, and their dysfunction is associated with several diseases. Understanding of the LLPS dynamics would...

journal article 2019

document

Shape and Size Control of Artificial Cells for Bottom-Up Biology

Fanalista, F. (author), Birnie, A.T.F. (author), Maan, R. (author), Burla, Federica (author), Charles usagé Charles-Carty, K.V. (author), Pawlik, G. (author), Deshpande, S.R. (author), Koenderink, Gijsje H. (author), Dogterom, A.M. (author), Dekker, C. (author)

Bottom-up biology is an expanding research field that aims to understand the mechanisms underlying biological processes via in vitro assembly of their essential components in synthetic cells. As encapsulation and controlled manipulation of these elements is a crucial step in the recreation of such cell-like objects, microfluidics is...

journal article 2019

document

Cytolinker Gas2L1 regulates axon morphology through microtubule-modulated actin stabilization

van de Willige, Dieudonnée (author), Hummel, Jessica J.A. (author), Alkemade, C. (author), Kahn, Olga I. (author), Au, Franco K.C. (author), Qi, Robert Z. (author), Dogterom, A.M. (author), Koenderink, G.H. (author), Hoogenraad, Casper C. (author), Akhmanova, Anna (author)

Crosstalk between the actin and microtubule cytoskeletons underlies cellular morphogenesis. Interactions between actin filaments and microtubules are particularly important for establishing the complex polarized morphology of neurons. Here, we characterized the neuronal function of growth arrest-specific 2-like 1 (Gas2L1), a protein that can...

journal article 2019

document

Minimal in vitro systems shed light on cell polarity

Vendel, K.J.A. (author), Tschirpke, S. (author), Shamsi, F. (author), Dogterom, A.M. (author), Laan, L. (author)

Cell polarity - the morphological and functional differentiation of cellular compartments in a directional manner - is required for processes such as orientation of cell division, directed cellular growth and motility. How the interplay of components within the complexity of a cell leads to cell polarity is still heavily debated. In this...

review 2019

document

Molecular determinants of the Ska-Ndc80 interaction and their influence on microtubule tracking and force-coupling

Huis In 't Veld, Pim J. (author), Volkov, V. (author), Stender, Isabelle D. (author), Musacchio, Andrea (author), Dogterom, A.M. (author)

Errorless chromosome segregation requires load-bearing attachments of the plus ends of spindle microtubules to chromosome structures named kinetochores. How these end-on kinetochore attachments are established following initial lateral contacts with the microtubule lattice is poorly understood. Two microtubule-binding complexes, the Ndc80 and...

journal article 2019

document

The depolymerase activity of MCAK shows a graded response to Aurora B kinase phosphorylation through allosteric regulation

McHugh, Toni (author), Zou, Juan (author), Volkov, V. (author), Bertin, Aurélie (author), Talapatra, Sandeep K. (author), Rappsilber, Juri (author), Dogterom, A.M. (author), Welburn, Julie P.I. (author)

Kinesin-13 motors regulate precise microtubule dynamics and limit microtubule length throughout metazoans by depolymerizing microtubule ends. Recently, the kinesin-13 motor family member MCAK (also known Kif2C) has been proposed to undergo large conformational changes during its catalytic cycle, as it switches from being in solution to being...

journal article 2019

document

CLASP Suppresses Microtubule Catastrophes through a Single TOG Domain

Aher, Amol (author), Kok, M.W.A. (author), Sharma, Ashwani (author), Rai, Ankit (author), Olieric, Natacha (author), Rodriguez-Garcia, Ruddi (author), Katrukha, Eugene A. (author), Weinert, Tobias (author), Olieric, Vincent (author), Kapitein, Lukas C. (author), Steinmetz, Michel O. (author), Dogterom, A.M. (author), Akhmanova, Anna (author)

The dynamic instability of microtubules plays a key role in controlling their organization and function, but the cellular mechanisms regulating this process are poorly understood. Here, we show that cytoplasmic linker-associated proteins (CLASPs) suppress transitions from microtubule growth to shortening, termed catastrophes, including those...

journal article 2018