EB1 interacts with outwardly curved and straight regions of the microtubule lattice.
EB1 is a microtubule plus-end tracking protein that recognizes GTP-tubulin dimers in microtubules and thus represents a unique probe to investigate the architecture of the GTP cap of growing microtubule ends. Here, we conjugated EB1 to gold nanoparticles (EB1-gold) and imaged by cryo-electron tomography its interaction with dynamic microtubules assembled in vitro from purified tubulin. EB1-gold forms comets at the ends of microtubules assembled in the presence of GTP, and interacts with the outer surface of curved and straight tubulin sheets as well as closed regions of the microtubule lattice. Microtubules assembled in the presence of GTP, different GTP analogues or cell extracts display similarly curved sheets at their growing ends, which gradually straighten as their protofilament number increases until they close into a tube. Together, our data provide unique structural information on the interaction of EB1 with growing microtubule ends. They further offer insights into the conformational changes that tubulin dimers undergo during microtubule assembly and the architecture of the GTP-cap region.
Institute of Genetics and Development of Rennes, UMR6290 CNRS, University of Rennes 1, Campus Universitaire de Beaulieu, 35042 Rennes Cédex, France.
Audrey
A
Guesdon
Institute of Genetics and Development of Rennes, UMR6290 CNRS, University of Rennes 1, Campus Universitaire de Beaulieu, 35042 Rennes Cédex, France.
Franck
F
Bazile
Laboratory of Biomolecular Research, Department of Biology and Chemistry, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.|Metabolic Engineering Group, Department of Microbiology and Genetics, University of Salamanca, Campus Miguel de Unamuno s/n, 37007 Salamanca, Spain.
Rubén M
RM
Buey
Cell Biology, Faculty of Science, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
Renu
R
Mohan
Institute of Genetics and Development of Rennes, UMR6290 CNRS, University of Rennes 1, Campus Universitaire de Beaulieu, 35042 Rennes Cédex, France.
Solange
S
Monier
Cell Biology, Faculty of Science, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
Ruddi Rodríguez
RR
García
Institute of Genetics and Development of Rennes, UMR6290 CNRS, University of Rennes 1, Campus Universitaire de Beaulieu, 35042 Rennes Cédex, France.
Morgane
M
Angevin
Institute of Genetics and Development of Rennes, UMR6290 CNRS, University of Rennes 1, Campus Universitaire de Beaulieu, 35042 Rennes Cédex, France.
Claire
C
Heichette
Institute of Biochemistry, Biocenter, and Cluster of Excellence-Macromolecular Complexes, Goethe-University Frankfurt, Max-von-Laue Strasse 9, 60438 Frankfurt am Main, Germany.
Ralph
R
Wieneke
Institute of Biochemistry, Biocenter, and Cluster of Excellence-Macromolecular Complexes, Goethe-University Frankfurt, Max-von-Laue Strasse 9, 60438 Frankfurt am Main, Germany.
Robert
0000-0002-0403-2160
R
Tampé
Institute of Genetics and Development of Rennes, UMR6290 CNRS, University of Rennes 1, Campus Universitaire de Beaulieu, 35042 Rennes Cédex, France.
Laurence
L
Duchesne
Cell Biology, Faculty of Science, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
Anna
0000-0002-9048-8614
A
Akhmanova
Laboratory of Biomolecular Research, Department of Biology and Chemistry, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
Michel O
MO
Steinmetz
Institute of Genetics and Development of Rennes, UMR6290 CNRS, University of Rennes 1, Campus Universitaire de Beaulieu, 35042 Rennes Cédex, France.|Microscopy Rennes Imaging Centre, and Biosit, UMS3480 CNRS, University of Rennes 1, Campus Santé de Villejean, 35043 Rennes Cédex, France.
Denis
D
Chrétien
D002460:Cell Line; D003599:Cytoskeleton / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000378:metabolism; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology; D011487:Protein Conformation; D014404:Tubulin / Q000378:metabolism
C500985:MAPRE1 protein, human; D008869:Microtubule-Associated Proteins; D014404:Tubulin; D006160:Guanosine Triphosphate