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Changes in seam number and location induce holes within microtubules assembled from porcine brain tubulin and in Xenopus egg cytoplasmic extracts.
Guyomar C
,
Bousquet C
,
Ku S
,
Heumann JM
,
Guilloux G
,
Gaillard N
,
Heichette C
,
Duchesne L
,
Steinmetz MO
,
Gibeaux R
,
Chrétien D
.
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Microtubules are tubes of about 25 nm in diameter that are critically involved in a variety of cellular functions, including motility, compartmentalization, and division. They are considered as pseudo-helical polymers whose constituent αβ-tubulin heterodimers share lateral homotypic interactions, except at one unique region called the seam. Here, we used a segmented sub-tomogram averaging strategy to reassess this paradigm and analyze the organization of the αβ-tubulin heterodimers in microtubules assembled from purified porcine brain tubulin in the presence of GTP and GMPCPP, and in Xenopus egg cytoplasmic extracts. We find that in almost all conditions, microtubules incorporate variable protofilament and/or tubulin subunit helical-start numbers, as well as variable numbers of seams. Strikingly, the seam number and location vary along individual microtubules, generating holes of one to a few subunits in size within their lattices. Together, our results reveal that the formation of mixed and discontinuous microtubule lattices is an intrinsic property of tubulin that requires the formation of unique lateral interactions without longitudinal ones. They further suggest that microtubule assembly is tightly regulated in a cytoplasmic environment.
Figure 1. Organization of tubulin within microtubules.
The αβ-tubulin heterodimers (α in cyan, β in yellow) alternate head-to-tail along protofilaments, 13 of which associate laterally to form the microtubule wall. (A) In the A-type lattice, the lateral contacts are made between heterotypic subunits (α-β, β-α) along the three-start helices. (B) In the B-type lattice, the lateral contacts are made between homotypic subunits (α-α, β-β), except at one unique region of the A-type called the seam. (C) Decoration of microtubules with kinesin-motor domains (orange) that bind to β-tubulin highlights the organization of the tubulin heterodimer within microtubules.
Akhmanova,
Tracking the ends: a dynamic protein network controls the fate of microtubule tips.
2008, Pubmed
Akhmanova,
Tracking the ends: a dynamic protein network controls the fate of microtubule tips.
2008,
Pubmed
Amos,
Arrangement of subunits in flagellar microtubules.
1974,
Pubmed
Bowne-Anderson,
Microtubule dynamic instability: a new model with coupled GTP hydrolysis and multistep catastrophe.
2013,
Pubmed
Bowne-Anderson,
Regulation of Microtubule Growth and Catastrophe: Unifying Theory and Experiment.
2015,
Pubmed
Brouhard,
XMAP215 is a processive microtubule polymerase.
2008,
Pubmed
,
Xenbase
Brouhard,
Dynamic instability 30 years later: complexities in microtubule growth and catastrophe.
2015,
Pubmed
Böhler,
The gamma-tubulin ring complex: Deciphering the molecular organization and assembly mechanism of a major vertebrate microtubule nucleator.
2021,
Pubmed
Carazo-Salas,
Generation of GTP-bound Ran by RCC1 is required for chromatin-induced mitotic spindle formation.
1999,
Pubmed
,
Xenbase
Castoldi,
Purification of brain tubulin through two cycles of polymerization-depolymerization in a high-molarity buffer.
2003,
Pubmed
Chaaban,
A microtubule bestiary: structural diversity in tubulin polymers.
2017,
Pubmed
Chen,
Eliminating effects of particle adsorption to the air/water interface in single-particle cryo-electron microscopy: Bacterial RNA polymerase and CHAPSO.
2019,
Pubmed
Chen,
Interaction of human erythrocyte catalase with air-water interface in cryoEM.
2022,
Pubmed
Chrétien,
New data on the microtubule surface lattice.
1991,
Pubmed
Chrétien,
Lattice defects in microtubules: protofilament numbers vary within individual microtubules.
1992,
Pubmed
,
Xenbase
Chrétien,
Microtubules switch occasionally into unfavorable configurations during elongation.
2000,
Pubmed
Cleary,
Molecular mechanisms underlying microtubule growth dynamics.
2021,
Pubmed
Crepeau,
Differences in alpha and beta polypeptide chains of tubulin resolved by electron microscopy with image reconstruction.
1978,
Pubmed
D'Imprima,
Protein denaturation at the air-water interface and how to prevent it.
2019,
Pubmed
Debs,
Dynamic and asymmetric fluctuations in the microtubule wall captured by high-resolution cryoelectron microscopy.
2020,
Pubmed
Dimitrov,
Detection of GTP-tubulin conformation in vivo reveals a role for GTP remnants in microtubule rescues.
2008,
Pubmed
Duchesne,
Robust ligand shells for biological applications of gold nanoparticles.
2008,
Pubmed
Edelstein,
Advanced methods of microscope control using μManager software.
2014,
Pubmed
Farmer,
XMAP215 promotes microtubule catastrophe by disrupting the growing microtubule end.
2021,
Pubmed
Foster,
A cryo-ET survey of microtubules and intracellular compartments in mammalian axons.
2022,
Pubmed
Gibeaux,
The Use of Cell-Free Xenopus Extracts to Investigate Cytoplasmic Events.
2019,
Pubmed
,
Xenbase
Good,
Preparation of Cellular Extracts from Xenopus Eggs and Embryos.
2018,
Pubmed
,
Xenbase
Guesdon,
Single versus dual-axis cryo-electron tomography of microtubules assembled in vitro: limits and perspectives.
2013,
Pubmed
Guesdon,
EB1 interacts with outwardly curved and straight regions of the microtubule lattice.
2016,
Pubmed
Hagen,
Implementation of a cryo-electron tomography tilt-scheme optimized for high resolution subtomogram averaging.
2017,
Pubmed
Helmke,
TPX2 levels modulate meiotic spindle size and architecture in Xenopus egg extracts.
2014,
Pubmed
,
Xenbase
Howes,
Structural differences between yeast and mammalian microtubules revealed by cryo-EM.
2017,
Pubmed
Hunyadi,
Mechanical stress induced mechanism of microtubule catastrophes.
2005,
Pubmed
Kikkawa,
Direct visualization of the microtubule lattice seam both in vitro and in vivo.
1994,
Pubmed
Klebl,
It started with a Cys: Spontaneous cysteine modification during cryo-EM grid preparation.
2022,
Pubmed
Kremer,
Computer visualization of three-dimensional image data using IMOD.
1996,
Pubmed
Li,
Effect of charge on protein preferred orientation at the air-water interface in cryo-electron microscopy.
2021,
Pubmed
Manka,
Microtubule structure by cryo-EM: snapshots of dynamic instability.
2018,
Pubmed
Mastronarde,
Dual-axis tomography: an approach with alignment methods that preserve resolution.
1997,
Pubmed
Maurer,
EBs recognize a nucleotide-dependent structural cap at growing microtubule ends.
2012,
Pubmed
McEwen,
Evidence for a mixed lattice in microtubules reassembled in vitro.
1980,
Pubmed
McIntosh,
Lattice structure of cytoplasmic microtubules in a cultured Mammalian cell.
2009,
Pubmed
McIntosh,
Microtubules grow by the addition of bent guanosine triphosphate tubulin to the tips of curved protofilaments.
2018,
Pubmed
Mitchison,
Dynamic instability of microtubule growth.
NULL,
Pubmed
Murray,
Cell cycle extracts.
1991,
Pubmed
Nicastro,
The molecular architecture of axonemes revealed by cryoelectron tomography.
2006,
Pubmed
Olieric,
Automated seamless DNA co-transformation cloning with direct expression vectors applying positive or negative insert selection.
2010,
Pubmed
Pantaloni,
Involvement of guanosine triphosphate (GTP) hydrolysis in the mechanism of tubulin polymerization: regulation of microtubule dynamics at steady state by a GTP cap.
1986,
Pubmed
Rai,
Lattice defects induced by microtubule-stabilizing agents exert a long-range effect on microtubule growth by promoting catastrophes.
2021,
Pubmed
Rothwell,
End-to-end annealing of microtubules in vitro.
1986,
Pubmed
Sawin,
Microtubule flux in mitosis is independent of chromosomes, centrosomes, and antiparallel microtubules.
1994,
Pubmed
,
Xenbase
Schaedel,
Lattice defects induce microtubule self-renewal.
2019,
Pubmed
Song,
Recombinant kinesin motor domain binds to beta-tubulin and decorates microtubules with a B surface lattice.
1993,
Pubmed
Song,
The anatomy of flagellar microtubules: polarity, seam, junctions, and lattice.
1995,
Pubmed
Sosa,
Three different approaches for calculating the three-dimensional structure of microtubules decorated with kinesin motor domains.
1997,
Pubmed
Théry,
Microtubule self-repair.
2021,
Pubmed
Vitre,
EB1 regulates microtubule dynamics and tubulin sheet closure in vitro.
2008,
Pubmed
Wade,
Cryoelectron microscopy of microtubules.
1993,
Pubmed
Zabeo,
A lumenal interrupted helix in human sperm tail microtubules.
2018,
Pubmed
Zhang,
Mechanistic Origin of Microtubule Dynamic Instability and Its Modulation by EB Proteins.
2015,
Pubmed
des Georges,
Mal3, the Schizosaccharomyces pombe homolog of EB1, changes the microtubule lattice.
2008,
Pubmed
