Structural differences between yeast and mammalian microtubules revealed by cryo-EM

Howes, Stuart C.; Geyer, Elisabeth A.; LaFrance, Benjamin; Zhang, Rui; Kellogg, Elizabeth H.; Westermann, Stefan; Rice, Luke M.; Nogales, Eva

doi:10.1083/jcb.201612195

Cited by 76 publications

(87 citation statements)

References 59 publications

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“…Interestingly, binding of Bim1 to yeast MTs was also distinct from the binding of EB3 to mammalian MTs. While the latter binds MTs with a tubulin dimer repeat, as most MAPs do, Bim1 binds yeast MTs with a tubulin monomer repeat, that is, double the stoichiometry [28]. A similar difference in binding pattern has also been previously observed for the Ndc80 kinetochore complex, which binds with the classical tubulin dimer repeat in C elegans, but with a tubulin monomer repeat in humans [7,20,21,29].…”

Section: Structure Of Mts From Alternative Sourcesmentioning

confidence: 59%

“…With the development of new purification strategies [24] and expression systems [25,26], it has recently become possible to study more pure tubulin samples from alternative sources. A number of recent structural studies have now used recombinant human tubulin [25,27] and purified yeast tubulin [28] (which is significantly simpler in terms of tubulin isoforms). The latter opens the possibility to carry out parallel in vitro and in vivo studies in the characterization of tubulin mutants, as well as to explore, given the evolutionary distance between yeast and mammalian tubulin, the conservation (or not) of different structural properties between these two MT systems.…”

Section: Structure Of Mts From Alternative Sourcesmentioning

confidence: 99%

“…Sequence differences around the lateral contacts may explain this observation. Both the number and nature of these sequence differences (a significant number of mammalian residues at the lateral contact correspond to serines in yeast tubulin) have the potential to alter slightly the geometry of the contacts, as well as their strength [28]. …”

Section: Structure Of Mts From Alternative Sourcesmentioning

confidence: 99%

“…More intriguingly, the conformational change in α-tubulin that gives rise to a lattice compaction upon GTP hydrolysis [12,17] (as observed by comparing mammalian MTs bound to the slowly hydrolysable GTP-analog GMPCPP versus those bound to GDP), is not seen in yeast MTs [28]. For yeast, both GMPCPP stabilized MTs and MT grown in the presence of GTP appear extended.…”

Section: Structure Of Mts From Alternative Sourcesmentioning

confidence: 99%

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Challenges and opportunities in the high-resolution cryo-EM visualization of microtubules and their binding partners

Nogales

Kellogg

2017

Current Opinion in Structural Biology

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As non-crystallizable polymers, microtubules have been the target of cryo-electron microscopy (cryo-EM) studies since the technique was first established. Over the years, image processing strategies have been developed that take care of the unique, pseudo-helical symmetry of the microtubule. With recent progress in data quality and data processing, cryo-EM reconstructions are now reaching resolutions that allow the generation of atomic models of microtubules and the factors that bind them. These include cellular partners that contribute to microtubule cellular functions, or small ligands that interfere with those functions in the treatment of cancer. The stage is set to generate a family portrait for all identified microtubule interacting proteins and to use cryo-EM as a drug development tool in the targeting of tubulin.

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Section: Structure Of Mts From Alternative Sourcesmentioning

confidence: 59%

Section: Structure Of Mts From Alternative Sourcesmentioning

confidence: 99%

Section: Structure Of Mts From Alternative Sourcesmentioning

confidence: 99%

Section: Structure Of Mts From Alternative Sourcesmentioning

confidence: 99%

See 2 more Smart Citations

Challenges and opportunities in the high-resolution cryo-EM visualization of microtubules and their binding partners

Nogales

Kellogg

2017

Current Opinion in Structural Biology

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“…Our previous purification of TBCD•ARL2 as a dimer that cannot bind GTP [33] is consistent with the binding of β-tubulin acting to change the nucleotide binding properties of the ARL2, specifically promoting GTP binding, which we hypothesize then allosterically alters β-tubulin structure into one that promotes binding to α-tubulin. Finally, there is steadily increasing evidence this work, our previous study [11], and a recent publication [87], that reveal important species differences in tubulins and microtubules that need to be considered when testing hypotheses regarding the complex folding or polymerization pathways.…”

Section: Discussionmentioning

confidence: 91%

Nucleotide Binding to ARL2 in the TBCD ∙ ARL2 ∙ β-Tubulin Complex Drives Conformational Changes in β-Tubulin

Francis

Goswami

Novick

et al. 2017

Journal of Molecular Biology

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Microtubules are highly dynamic tubulin polymers that are required for a variety of cellular functions. Despite the importance of a cellular population of tubulin dimers, we have incomplete information about the mechanisms involved in the biogenesis of αβ-tubulin heterodimers. In addition to prefoldin and the TCP-1 Ring Complex, five tubulin-specific chaperones, termed cofactors A-E (TBCA-E), and GTP are required for the folding of α- and β-tubulin subunits and assembly into heterodimers. We recently described the purification of a novel trimer, TBCD•ARL2•β-tubulin. Here, we employed hydrogen/deuterium exchange coupled with mass spectrometry to explore the dynamics of each of the proteins in the trimer. Addition of guanine nucleotides resulted in changes in the solvent accessibility of regions of each protein that led to predictions about each’s role in tubulin folding. Initial testing of that model confirmed that it is ARL2, and not β-tubulin, that exchanges GTP in the trimer. Comparisons of the dynamics of ARL2 monomer to ARL2 in the trimer suggested that its protein interactions were comparable to those of a canonical GTPase with an effector. This was supported by the use of nucleotide binding assays that revealed an increase in the affinity for GTP by ARL2 in the trimer. We conclude that the TBCD•ARL2•β-tubulin complex represents a functional intermediate in the β-tubulin folding pathway whose activity is regulated by the cycling of nucleotides on ARL2. The co-purification of guanine nucleotide on the β-tubulin in the trimer is also shown, with implications to modeling the pathway.

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Non‐Equilibrium Assembly of Atomically‐Precise Copper Nanoclusters

Zhao,

Xu,

et al. 2024

Advanced Materials

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Accurate structure control in dissipative assemblies is vital for precise biological functions. However, the accuracy and functionality of current artificial dissipative assembly are far from this objective. Herein, we introduce a novel approach by harnessing complex chemical reaction networks (CRN) rooted in coordination chemistry to create well‐defined dissipative assemblies. We designed atomically‐precise Cu nanocluster (CuNCs), specifically Cu11(μ9‐Cl)(μ3‐Cl)3L6Cl clusters (L = 4‐methyl‐piperazine‐1‐carbodithioate). Cu(I)‐ligand ratio change and dynamic Cu(I)‐Cu(I) metallophilic/coordination interactions enable the reorganization of CuNCs into metastable CuL2, finally converting into the equilibrium [CuL·Y]Cl complexes (Y = MeCN or H2O) via Cu(I) oxidation/reorganization and ligand exchange process. Upon adding fuels (ascorbic acid, AA), the system goes further dissipative cycles. We observed that the encapsulated/bridging halide ions exert a subtle influence on the optical properties of CuNCs and topological changes of polymeric networks when integrating CuNCs as crosslink sites. CuNCs duration and switch period could be controlled by varying the ions, AA concentration, O2 pressure and pH. The unique Cu(I)‐Cu(I) metallophilic and coordination interactions provide a versatile toolbox for designing delicate life‐like materials, paving the way for tailored dissipative assemblies with precise structures and functionalities. Furthermore, these CuNCs can be employed as modular units within polymers for materials mechanics or functionalization studies, expanding their potential applications.This article is protected by copyright. All rights reserved

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Structural differences between yeast and mammalian microtubules revealed by cryo-EM

Abstract: Yeast MTs do not appear to undergo the lattice compaction seen in mammalian MTs upon GTP hydrolysis. Binding of the +TIP Bim1, both between and within αβ-tubulin dimers, causes compaction of yeast MTs and their rapid disassembly.

Cited by 76 publications

References 59 publications

Challenges and opportunities in the high-resolution cryo-EM visualization of microtubules and their binding partners

Challenges and opportunities in the high-resolution cryo-EM visualization of microtubules and their binding partners

Nucleotide Binding to ARL2 in the TBCD ∙ ARL2 ∙ β-Tubulin Complex Drives Conformational Changes in β-Tubulin

Non‐Equilibrium Assembly of Atomically‐Precise Copper Nanoclusters

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