Project

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Protein interactions regulating the microtubule cytoskeleton

English title Protein interactions regulating the microtubule cytoskeleton
Applicant Steinmetz Michel
Number 138659
Funding scheme Project funding (Div. I-III)
Research institution Paul Scherrer Institut
Institution of higher education Paul Scherrer Institute - PSI
Main discipline Biochemistry
Start/End 01.01.2012 - 31.12.2015
Approved amount 863'000.00
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All Disciplines (2)

Discipline
Biochemistry
Biophysics

Keywords (4)

microtubule cytoskeleton; microtubule plus-end tracking proteins; centriolar proteins; microtubule-modifying enzymes

Lay Summary (English)

Lead
Protein interactions regulating the microtubule cytoskeleton
Lay summary

The microtubule cytoskeleton controls fundamental cellular processes including mitosis, cell polarity, and intracellular transport. Because of its importance in all these vital processes, microtubule networks are further implicated in several human pathological conditions including cancer. In cells, microtubule associated proteins interact with the microtubule cytoskeleton to regulate its diverse activities both spatially and temporally during the cell cycle. To understand in detail the molecular mechanisms of these proteins and their interplay is of considerable importance both in health and disease and represents a major challenge. In this proposal we plan to work on three subprojects related to the microtubule cytoskeleton:

1. Structure-function relationship of microtubule plus-end tracking proteins

2. Mechanisms of centriole formation

3. Mechanisms of tubulin modifying enzymes

The aim of the project is to provide detailed quantitative (equilibrium, kinetic), structural (high and medium resolution) and functional information for understanding the proteins and their interaction networks involved in the three subprojects at the molecular level. To reach this goal we plan to use X-ray crystallography in combination with biochemical and biophysical methods. In collaboration, we furthermore will use cell biological methods to test emerging concepts derived from in vitro experiments directly in different cellular systems. Our proposed work will provide novel insights into the molecular mechanisms of several key proteins centrally involved in the regulation of the microtubule cytoskeleton.

Direct link to Lay Summary Last update: 12.11.2012

Responsible applicant and co-applicants

Employees

Publications

Publication
Antivascular and antitumor properties of the tubulin-binding chalcone TUB091
Canela María-Dolores, Noppen Sam, Bueno Oskía, Prota Andrea E., Bargsten Katja, Sáez-Calvo Gonzalo, Jimeno María-Luisa, Benkheil Mohammed, Ribatti Domenico, Velázquez Sonsoles, Camarasa María-José, Díaz J. Fernando, Steinmetz Michel O., Priego Eva-María, Pérez-Pérez María-Jesús, Liekens Sandra (2017), Antivascular and antitumor properties of the tubulin-binding chalcone TUB091, in Oncotarget, 1.
Biophysical and Structural Characterization of the Centriolar Protein Cep104 Interaction Network
Rezabkova Lenka, Kraatz Sebastian H. W., Akhmanova Anna, Steinmetz Michel O., Kammerer Richard A. (2016), Biophysical and Structural Characterization of the Centriolar Protein Cep104 Interaction Network, in Journal of Biological Chemistry, 291(35), 18496-18504.
Centriolar CPAP/SAS-4 Imparts Slow Processive Microtubule Growth
Sharma Ashwani, Aher Amol, Dynes Nicola J., Frey Daniel, Katrukha Eugene A., Jaussi Rolf, Grigoriev Ilya, Croisier Marie, Kammerer Richard A., Akhmanova Anna, Gönczy Pierre, Steinmetz Michel O. (2016), Centriolar CPAP/SAS-4 Imparts Slow Processive Microtubule Growth, in Developmental Cell, 37(4), 362-376.
Crystal Structures of the Human Doublecortin C- and N-terminal Domains in Complex with Specific Antibodies
Burger Dominique, Stihle Martine, Sharma Ashwani, Di Lello Paola, Benz Jörg, D'Arcy Brigitte, Debulpaep Maja, Fry David, Huber Walter, Kremer Thomas, Laeremans Toon, Matile Hugues, Ross Alfred, Rufer Arne C., Schoch Guillaume, Steinmetz Michel O., Steyaert Jan, Rudolph Markus G., Thoma Ralf, Ruf Armin (2016), Crystal Structures of the Human Doublecortin C- and N-terminal Domains in Complex with Specific Antibodies, in Journal of Biological Chemistry, 291(31), 16292-16306.
Data-collection strategy for challenging native SAD phasing
Olieric Vincent, Weinert Tobias, Finke Aaron D., Anders Carolin, Li Dianfan, Olieric Natacha, Borca Camelia N., Steinmetz Michel O., Caffrey Martin, Jinek Martin, Wang Meitian (2016), Data-collection strategy for challenging native SAD phasing, in Acta Crystallographica Section D Structural Biology, 72(3), 421-429.
EB1 interacts with outwardly curved and straight regions of the microtubule lattice
Guesdon Audrey, Bazile Franck, Buey Rubén M., Mohan Renu, Monier Solange, García Ruddi Rodríguez, Angevin Morgane, Heichette Claire, Wieneke Ralph, Tampé Robert, Duchesne Laurence, Akhmanova Anna, Steinmetz Michel O., Chrétien Denis (2016), EB1 interacts with outwardly curved and straight regions of the microtubule lattice, in Nature Cell Biology, 18(10), 1102-1108.
Kinesin-Binding Protein Controls Microtubule Dynamics and Cargo Trafficking by Regulating Kinesin Motor Activity
Kevenaar Josta T., Bianchi Sarah, van Spronsen Myrrhe, Olieric Natacha, Lipka Joanna, Frias Cátia P., Mikhaylova Marina, Harterink Martin, Keijzer Nanda, Wulf Phebe S., Hilbert Manuel, Kapitein Lukas C., de Graaff Esther, Ahkmanova Anna, Steinmetz Michel O., Hoogenraad Casper C. (2016), Kinesin-Binding Protein Controls Microtubule Dynamics and Cargo Trafficking by Regulating Kinesin Motor Activity, in Current Biology, 26(7), 849-861.
Molecular basis of Kar9-Bim1 complex function during mating and spindle positioning
Manatschal Cristina, Farcas Ana-Maria, Degen Miriam Steiner, Bayer Mathias, Kumar Anil, Landgraf Christiane, Volkmer Rudolf, Barral Yves, Steinmetz Michel O. (2016), Molecular basis of Kar9-Bim1 complex function during mating and spindle positioning, in Molecular Biology of the Cell, 27(23), 3729-3745.
Pironetin Binds Covalently to αCys316 and Perturbs a Major Loop and Helix of α-Tubulin to Inhibit Microtubule Formation
Prota Andrea E., Setter Jocelyn, Waight Andrew B., Bargsten Katja, Murga Juan, Diaz José Fernando, Steinmetz Michel O. (2016), Pironetin Binds Covalently to αCys316 and Perturbs a Major Loop and Helix of α-Tubulin to Inhibit Microtubule Formation, in Journal of Molecular Biology, 428(15), 2981-2988.
SAS-6 engineering reveals interdependence between cartwheel and microtubules in determining centriole architecture
Hilbert Manuel, Noga Akira, Frey Daniel, Hamel Virginie, Guichard Paul, Kraatz Sebastian H. W., Pfreundschuh Moritz, Hosner Sarah, Flückiger Isabelle, Jaussi Rolf, Wieser Mara M., Thieltges Katherine M., Deupi Xavier, Müller Daniel J., Kammerer Richard A., Gönczy Pierre, Hirono Masafumi, Steinmetz Michel O. (2016), SAS-6 engineering reveals interdependence between cartwheel and microtubules in determining centriole architecture, in Nature Cell Biology, 18(4), 393-403.
Structural basis for misregulation of kinesin KIF21A autoinhibition by CFEOM1 disease mutations
Bianchi Sarah, van Riel Wilhelmina E., Kraatz Sebastian H. W., Olieric Natacha, Frey Daniel, Katrukha Eugene A., Jaussi Rolf, Missimer John, Grigoriev Ilya, Olieric Vincent, Benoit Roger M., Steinmetz Michel O., Akhmanova Anna, Kammerer Richard A. (2016), Structural basis for misregulation of kinesin KIF21A autoinhibition by CFEOM1 disease mutations, in Scientific Reports, 6(1), 1.
Structural Basis of Microtubule Destabilization by Potent Auristatin Anti-Mitotics
Waight Andrew B., Bargsten Katja, Doronina Svetlana, Steinmetz Michel O., Sussman Django, Prota Andrea E. (2016), Structural Basis of Microtubule Destabilization by Potent Auristatin Anti-Mitotics, in PLOS ONE, 11(8), e0160890-e0160890.
Structural Determinants of the Dictyostatin Chemotype for Tubulin Binding Affinity and Antitumor Activity Against Taxane- and Epothilone-Resistant Cancer Cells
Trigili Chiara, Barasoain Isabel, Sánchez-Murcia Pedro A., Bargsten Katja, Redondo-Horcajo Mariano, Nogales Aurora, Gardner Nicola M., Meyer Arndt, Naylor Guy J., Gómez-Rubio Elena, Gago Federico, Steinmetz Michel O., Paterson Ian, Prota Andrea E., Díaz J. Fernando (2016), Structural Determinants of the Dictyostatin Chemotype for Tubulin Binding Affinity and Antitumor Activity Against Taxane- and Epothilone-Resistant Cancer Cells, in ACS Omega, 1(6), 1192-1204.
Termination of Protofilament Elongation by Eribulin Induces Lattice Defects that Promote Microtubule Catastrophes
Doodhi Harinath, Prota Andrea E., Rodríguez-García Ruddi, Xiao Hui, Custar Daniel W., Bargsten Katja, Katrukha Eugene A., Hilbert Manuel, Hua Shasha, Jiang Kai, Grigoriev Ilya, Yang Chia-Ping H., Cox David, Horwitz Susan Band, Kapitein Lukas C., Akhmanova Anna, Steinmetz Michel O. (2016), Termination of Protofilament Elongation by Eribulin Induces Lattice Defects that Promote Microtubule Catastrophes, in Current Biology, 26(13), 1713-1721.
The Human Centriolar Protein CEP135 Contains a Two-Stranded Coiled-Coil Domain Critical for Microtubule Binding
Kraatz Sebastian, Guichard Paul, Obbineni Jagan M., Olieric Natacha, Hatzopoulos Georgios N., Hilbert Manuel, Sen Indrani, Missimer John, Gönczy Pierre, Steinmetz Michel O. (2016), The Human Centriolar Protein CEP135 Contains a Two-Stranded Coiled-Coil Domain Critical for Microtubule Binding, in Structure, 24(8), 1358-1371.
Control of microtubule organization and dynamics: two ends in the limelight
Akhmanova Anna, Steinmetz Michel O. (2015), Control of microtubule organization and dynamics: two ends in the limelight, in NATURE REVIEWS MOLECULAR CELL BIOLOGY, 16(12), 711-726.
Fast native-SAD phasing for routine macromolecular structure determination (vol 12, pg 131, 2015)
Weinert Tobias, Olieric Vincent, Waltersperger Sandro, Panepucci Ezequiel, Chen Lirong, Zhang Hua, Zhou Dayong, Rose John, Ebihara Akio, Kuramitsu Seiki, Li Dianfan, Howe Nicole, Schnapp Gisela, Pautsch Alexander, Bargsten Katja, Prota Andrea E., Surana Parag, Kottur Jithesh, Nair Deepak T., Basilico Federica, Cecatiello Valentina, Pasqualato Sebastiano, Boland Andreas, Weichenrieder Oliver, Wang Bi-Cheng (2015), Fast native-SAD phasing for routine macromolecular structure determination (vol 12, pg 131, 2015), in NATURE METHODS, 12(7), 692-692.
Optochemistry to control the microtubule cytoskeleton
Janke Carsten, Steinmetz Michel O. (2015), Optochemistry to control the microtubule cytoskeleton, in EMBO JOURNAL, 34(16), 2114-2116.
A new tubulin-binding site and pharmacophore for microtubule-destabilizing anticancer drugs
Prota Andrea E., Bargsten Katja, Fernando Diaz J., Marsh May, Cuevas Carmen, Liniger Marc, Neuhaus Christian, Andreu Jose M., Altmann Karl-Heinz, Steinmetz Michel O. (2014), A new tubulin-binding site and pharmacophore for microtubule-destabilizing anticancer drugs, in PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 111(38), 13817-13821.
Localizing Chemical Groups while Imaging Single Native Proteins by High-Resolution Atomic Force Microscopy
Pfreundschuh M., Alsteens D., Hilbert M., Steinmetz M.O., Muller D.J. (2014), Localizing Chemical Groups while Imaging Single Native Proteins by High-Resolution Atomic Force Microscopy, in Nano. Lett., 14, 2957-2964.
Reconstitution of a hierarchical plus TIP interaction network controlling microtubule end tracking of dynein
Duellberg Christian, Trokter Martina, Jha Rupam, Sen Indrani, Steinmetz Michel O., Surrey Thomas (2014), Reconstitution of a hierarchical plus TIP interaction network controlling microtubule end tracking of dynein, in NATURE CELL BIOLOGY, 16(8), 804-804.
Structural basis for recognition of synaptic vesicle protein 2C by botulinum neurotoxin A
Benoit Roger M., Frey Daniel, Hilbert Manuel, Kevenaar Josta T., Wieser Mara M., Stirnimann Christian U., McMillan David, Ceska Tom, Lebon Florence, Jaussi Rolf, Steinmetz Michel O., Schertler Gebhard F. X., Hoogenraad Casper C., Capitani Guido, Kammerer Richard A. (2014), Structural basis for recognition of synaptic vesicle protein 2C by botulinum neurotoxin A, in NATURE, 505(7481), 108-108.
Structural Basis of Microtubule Stabilization by Laulimalide and Peloruside A
Prota Andrea E., Bargsten Katja, Northcote Peter T., Marsh May, Altmann Karl-Heinz, Miller John H., Fernando Diaz Jose, Steinmetz Michel O. (2014), Structural Basis of Microtubule Stabilization by Laulimalide and Peloruside A, in ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 53(6), 1621-1625.
Caenorhabditis elegans centriolar protein SAS-6 forms a spiral that is consistent with imparting a ninefold symmetry
Hilbert Manuel, Erat Michele C., Hachet Virginie, Guichard Paul, Blank Iris D., Flueckiger Isabelle, Slater Leanne, Lowe Edward D., Hatzopoulos Georgios N., Steinmetz Michel O., Goenczy Pierre, Vakonakis Ioannis (2013), Caenorhabditis elegans centriolar protein SAS-6 forms a spiral that is consistent with imparting a ninefold symmetry, in PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 110(28), 11373-11378.
End Binding Proteins Are Obligatory Dimers
Sen Indrani, Veprintsev Dmitry, Akhmanova Anna, Steinmetz Michel O. (2013), End Binding Proteins Are Obligatory Dimers, in PLOS ONE, 8(9), 1-6.
End-binding proteins sensitize microtubules to the action of microtubule-targeting agents
Mohan Renu, Katrukha Eugene A., Doodhi Harinath, Smal Ihor, Meijering Erik, Kapitein Lukas C., Steinmetz Michel O., Akhmanova Anna (2013), End-binding proteins sensitize microtubules to the action of microtubule-targeting agents, in PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 110(22), 8900-8905.
Molecular Mechanism of Action of Microtubule-Stabilizing Anticancer Agents
Prota Andrea E., Bargsten Katja, Zurwerra Didier, Field Jessica J., Fernando Diaz Jose, Altmann Karl-Heinz, Steinmetz Michel O. (2013), Molecular Mechanism of Action of Microtubule-Stabilizing Anticancer Agents, in SCIENCE, 339(6119), 587-590.
Structural Basis for the Oligomerization-State Switch from a Dimer to a Trimer of an Engineered Cortexillin-1 Coiled-Coil Variant
Bjelic Sasa, Wieser Mara, Frey Daniel, Stirnimann Christian U., Chance Mark R., Jaussi Rolf, Steinmetz Michel O., Kammerer Richard A. (2013), Structural Basis for the Oligomerization-State Switch from a Dimer to a Trimer of an Engineered Cortexillin-1 Coiled-Coil Variant, in PLOS ONE, 8(5), 1-6.
Structural basis of tubulin tyrosination by tubulin tyrosine ligase
Prota Andrea E., Magiera Maria M., Kuijpers Marijn, Bargsten Katja, Frey Daniel, Wieser Mara, Jaussi Rolf, Hoogenraad Casper C., Kammerer Richard A., Janke Carsten, Steinmetz Michel O. (2013), Structural basis of tubulin tyrosination by tubulin tyrosine ligase, in JOURNAL OF CELL BIOLOGY, 200(3), 259-270.
Cooperative stabilization of microtubule dynamics by EB1 and CLIP-170 involves displacement of stably bound P(i) at microtubule ends
Lopus M., Manatschal C., Buey R.M., Bjelic S., Miller H.P., Steinmetz M.O., Wilson L. (2012), Cooperative stabilization of microtubule dynamics by EB1 and CLIP-170 involves displacement of stably bound P(i) at microtubule ends, in Biochemistry, 51, 3021-3030.
Interaction of mammalian end binding proteins with CAP-Gly domains of CLIP-170 and p150(glued)
Bjelic S., De Groot C.O., Scharer M.A., Jaussi R., Bargsten K., Salzmann M., Frey D., Capitani G., Kammerer R.A., Steinmetz M.O. (2012), Interaction of mammalian end binding proteins with CAP-Gly domains of CLIP-170 and p150(glued), in J. Struct. Biol., 177, 160-167.
Sequence determinants of a microtubule tip localization signal (MtLS)
Buey R.M., Sen I., Kortt O., Mohan R., Gfeller D., Veprintsev D., Kretzschmar I., Scheuermann J., Neri D., Zoete V., Michielin O., de Pereda J.M., Akhmanova A., Volkmer R., Steinmetz M.O. (2012), Sequence determinants of a microtubule tip localization signal (MtLS), in J. Biol. Chem., 287(34), 28227-28242.
Tyrosine-dependent capture of CAP-Gly domain-containing proteins in complex mixture by EB1 C-terminal peptidic probes
Calligaris D., Manatschal C., Marcellin M., Villard C., Monsarrat B., Burlet-Schiltz O., Steinmetz M.O., Braguer D., Lafitte D., Verdier-Pinard P. (2012), Tyrosine-dependent capture of CAP-Gly domain-containing proteins in complex mixture by EB1 C-terminal peptidic probes, in J. Proteomics, 75, 3605-3616.

Collaboration

Group / person Country
Types of collaboration
Pierre Gönczy, EPFL Lausanne Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Yves Barral, ETH Zürich Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
Carsten Janke, Institut Curie, Paris France (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Anna Akhmanova, Erasmus Medical Center, Rotterdam Netherlands (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication

Scientific events



Self-organised

Title Date Place
EMBO Conference Series: Microtubules: Structure, Regulation and Functions 23.05.2012 Heidelberg, Germany

Associated projects

Number Title Start Funding scheme
166608 Control of Tubulin Structure and Function by Accessory Proteins and Drugs 01.04.2016 Project funding (Div. I-III)
125463 An integrated multidisciplinary approach towards a molecular understanding of centrosome duplication 01.01.2010 Sinergia
122545 Mechanisms of dynamic +TIP interactions 01.01.2009 Project funding (Div. I-III)

Abstract

The microtubule cytoskeleton controls fundamental cellular processes including mitosis, cell division, and intracellular transport. Because of its importance in all these vital processes, microtubule networks are further implicated in several human pathological conditions including cancer. In cells, microtubule associated proteins interact with the microtubule cytoskeleton to regulate its diverse activities both spatially and temporally during the cell cycle. To understand in detail the molecular mechanisms of these proteins and their interplay is of considerable importance both in health and disease and represents a major challenge. In this proposal we plan to work on three subprojects related to the microtubule cytoskeleton:1. Structure-function relationship of microtubule plus-end tracking proteins2. Mechanisms of centriole formation3. Mechanisms of tubulin modifying enzymesThe aim of the proposal is to provide detailed quantitative (equilibrium, kinetic), structural (high and medium resolution) and functional information for understanding the proteins and their interaction networks involved in the three subprojects at the molecular level. To reach this goal we plan to use X-ray crystallography in combination with biochemical and biophysical methods. In collaboration, we furthermore will use cell biological methods to test emerging concepts derived from in vitro experiments directly in different cellular systems. Our proposed work will provide novel insights into the molecular mechanisms of several key proteins centrally involved in the regulation of the microtubule cytoskeleton.
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