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Grant support

We thank Andrea Nans at the Structural Biology Science Technology Platform (STP) for support on the Titan Krios, Raffa Carzaniga at the Electron Microscopy STP of the Francis Crick Institute for support on the Tecnai G2 Spirit electron microscope, Nicholas I. Cade for fluorescence microscopy support, and Andrew Purkiss and Phil Walker for computational support. We thank the Cell Services, the flow cytometry, and the mass spectrometry proteomics STPs of the Francis Crick Institute for producing large cell cultures and for cell sorting and protein identification support. This work was supported by the Francis Crick Institute, which receives its core funding from Cancer Research UK (FC001163 and FC0010065), the UK Medical Research Council (FC001163 and FC0010065), and the Wellcome Trust (FC001163 and FC0010065) to T.S. and A.C. The Wellcome Centre for Cell Biology is supported by core funding from the Wellcome Trust (203149). J. Rappsilber is funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany's Excellence Strategy - EXC 2008 -390540038 -UniSysCat and 329673113. J. Roostalu. was supported by a Sir Henry Wellcome Postdoctoral Fellowship (100145/Z/12/Z) and M.A.C. is supported by a Marie Sk1odowskaCurie Postdoctoral Fellowship (agreement no. 845939). T.S. acknowledges support from the European Research Council (Advanced Grant, project 323042). A.C. receives funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation program (grant agreement no. 820102). T.C., J.A., J.W.M., and T.S. acknowledge also the support of the Spanish Ministry of Economy, Industry and Competitiveness to the CRG-EMBL partnership, the Centro de Excelencia Severo Ochoa and the CERCA Programme of the Generalitat de Catalunya.

Analysis of institutional authors

Consolati, TAuthorAsthana, JAuthorLim, WmAuthorSurrey, TCorresponding Author

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July 6, 2020
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Microtubule Nucleation Properties of Single Human gamma TuRCs Explained by Their Cryo-EM Structure

Publicated to:Developmental Cell. 53 (5): 603-+ - 2020-06-08 53(5), DOI: 10.1016/j.devcel.2020.04.019

Authors: Consolati, T; Locke, J; Roostalu, J; Chen, ZA; Gannon, J; Asthana, J; Lim, WM; Martino, F; Cvetkovic, MA; Rappsilber, J; Costa, A; Surrey, T

Affiliations

BIST, Ctr Genom Regulat CRG, Dr Aiguader 88, Barcelona 08003, Spain - Author
Francis Crick Inst, 1 Midland Rd, London NW1 1AT, England - Author
ICREA, Passeig de Lluis Co 23, Barcelona 08010, Spain - Author
Tech Univ Berlin, Inst Biotechnol, Bioanalyt, Berlin, Germany - Author
Univ Edinburgh, Wellcome Ctr Cell Biol, Edinburgh, Midlothian, Scotland - Author
Wellcome Trust Res Labs, London, England - Author
Wellcome Trust Res Labs, London, EnglandFrancis Crick Inst, 1 Midland Rd, London NW1 1AT, England - Author
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Abstract

The gamma-tubulin ring complex (gamma TuRC) is the major microtubule nucleator in cells. The mechanism of its regulation is not understood. We purified human gamma TuRC and measured its nucleation properties in a total internal reflection fluorescence (TIRF) microscopy-based real-time nucleation assay. We find that gamma TuRC stably caps the minus ends of microtubules that it nucleates stochastically. Nucleation is inefficient compared with microtubule elongation. The 4 angstrom resolution cryoelectron microscopy (cryo-EM) structure of gamma TuRC, combined with crosslinking mass spectrometry analysis, reveals an asymmetric conformation with only part of the complex in a "closed" conformation matching the microtubule geometry. Actin in the core of the complex, and MZT2 at the outer perimeter of the closed part of gamma TuRC appear to stabilize the closed conformation. The opposite side of gamma TuRC is in an "open," nucleation-incompetent conformation, leading to a structural asymmetry explaining the low nucleation efficiency of purified human gamma TuRC. Our data suggest possible regulatory mechanisms for microtubule nucleation by gamma TuRC closure.

Keywords

?-tubulin ring complexactinchtogclmscryo-electron microscopycrystal-structuredynamicsimportin-alphakineticsmicrotubule nucleationmzt2proteinringtirf microscopytpx2tubulin complexesxmap215γ-tubulin ring complex?-tubulin ring complex?turc structureActinArticleBiotinylationCarboxy terminal sequenceChromosome segregationChtogClmsComplex formationControlled studyCryo-electron microscopyCryoelectron microscopyFemaleGamma tubulinGamma tubulin ring complexHumanHuman cellLiquid chromatography-mass spectrometryMicrotubuleMicrotubule associated proteinMicrotubule nucleationMzt2Mzt2 proteinNonhumanPolyacrylamide gel electrophoresisPriority journalProtein conformationProtein cross linkingProtein purificationProtein stabilityProtein structureRegulatory mechanismTirf microscopyTotal internal reflection fluorescence microscopyTpx2Tubulin polymerization assayUnclassified drugWestern blottingΓturc structure

Quality index

Bibliometric impact. Analysis of the contribution and dissemination channel

The work has been published in the journal Developmental Cell due to its progression and the good impact it has achieved in recent years, according to the agency WoS (JCR), it has become a reference in its field. In the year of publication of the work, 2020, it was in position 2/41, thus managing to position itself as a Q1 (Primer Cuartil), in the category Developmental Biology. Notably, the journal is positioned above the 90th percentile.

From a relative perspective, and based on the normalized impact indicator calculated from World Citations provided by WoS (ESI, Clarivate), it yields a value for the citation normalization relative to the expected citation rate of: 2.39. This indicates that, compared to works in the same discipline and in the same year of publication, it ranks as a work cited above average. (source consulted: ESI Nov 14, 2024)

This information is reinforced by other indicators of the same type, which, although dynamic over time and dependent on the set of average global citations at the time of their calculation, consistently position the work at some point among the top 50% most cited in its field:

  • Weighted Average of Normalized Impact by the Scopus agency: 1.77 (source consulted: FECYT Feb 2024)
  • Field Citation Ratio (FCR) from Dimensions: 14.92 (source consulted: Dimensions Jul 2025)

Specifically, and according to different indexing agencies, this work has accumulated citations as of 2025-07-08, the following number of citations:

  • WoS: 82
  • Scopus: 41
  • Europe PMC: 56

Impact and social visibility

From the perspective of influence or social adoption, and based on metrics associated with mentions and interactions provided by agencies specializing in calculating the so-called "Alternative or Social Metrics," we can highlight as of 2025-07-08:

  • The use, from an academic perspective evidenced by the Altmetric agency indicator referring to aggregations made by the personal bibliographic manager Mendeley, gives us a total of: 98.
  • The use of this contribution in bookmarks, code forks, additions to favorite lists for recurrent reading, as well as general views, indicates that someone is using the publication as a basis for their current work. This may be a notable indicator of future more formal and academic citations. This claim is supported by the result of the "Capture" indicator, which yields a total of: 103 (PlumX).

With a more dissemination-oriented intent and targeting more general audiences, we can observe other more global scores such as:

  • The Total Score from Altmetric: 3.5.
  • The number of mentions on the social network X (formerly Twitter): 5 (Altmetric).

It is essential to present evidence supporting full alignment with institutional principles and guidelines on Open Science and the Conservation and Dissemination of Intellectual Heritage. A clear example of this is:

  • The work has been submitted to a journal whose editorial policy allows open Open Access publication.

Leadership analysis of institutional authors

This work has been carried out with international collaboration, specifically with researchers from: Germany; United Kingdom.

There is a significant leadership presence as some of the institution’s authors appear as the first or last signer, detailed as follows: First Author (CONSOLATI, TANJA) and Last Author (SURREY, THOMAS).

the author responsible for correspondence tasks has been SURREY, THOMAS.