ArticleMolecular biology of the cell2002
Reconstitution and characterization of budding yeast gamma-tubulin complex.
Article in Molecular biology of the cell, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 48 papers.
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Who cites it
48 citing papers in PubMed, 76 citations in OpenAlex.
- The TOG protein Stu2 is regulated by acetylation.PLoS genetics · 2022Article
- B1-type cyclins control microtubule organization during cell division in Arabidopsis.EMBO reports · 2022Article
- Molecular insight into how γ-TuRC makes microtubules.Journal of cell science · 2021Review
- CM1-driven assembly and activation of yeast γ-tubulin small complex underlies microtubule nucleation.eLife · 2021Article
- Biochemical reconstitutions reveal principles of human γ-TuRC assembly and function.The Journal of cell biology · 2021Article
- Yeast pericentrin/Spc110 contains multiple domains required for tethering the γ-tubulin complex to the centrosome.Molecular biology of the cell · 2020Article
- Article
- Reconstitution of Microtubule Nucleation In Vitro Reveals Novel Roles for Mzt1.Current biology : CB · 2019Article
- The XMAP215 Ortholog Alp14 Promotes Microtubule Nucleation in Fission Yeast.Current biology : CB · 2018Article
- Article
- Stimulation of microtubule-based transport by nucleation of microtubules on pigment granules.Molecular biology of the cell · 2017Article
- Metaphase Spindle Assembly.Biology · 2017Review
- MOZART1 and γ-tubulin complex receptors are both required to turn γ-TuSC into an active microtubule nucleation template.The Journal of cell biology · 2016Article
- Higher-order oligomerization of Spc110p drives γ-tubulin ring complex assembly.Molecular biology of the cell · 2016Article
- The molecular architecture of the Dam1 kinetochore complex is defined by cross-linking based structural modelling.Nature communications · 2015Article
- γ-Tubulin complex in Trypanosoma brucei: molecular composition, subunit interdependence and requirement for axonemal central pair protein assembly.Molecular microbiology · 2015Article
- Mto2 multisite phosphorylation inactivates non-spindle microtubule nucleation complexes during mitosis.Nature communications · 2015Article
- Microtubule-associated proteins control the kinetics of microtubule nucleation.Nature cell biology · 2015Article
- Ring closure activates yeast γTuRC for species-specific microtubule nucleation.Nature structural & molecular biology · 2015Article
- Lessons from yeast: the spindle pole body and the centrosome.Philosophical transactions of the Royal Society of London. Series B, Biological sciences · 2014Review
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Authors and funding
5 authors at 2 institutions in 1 country.
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Abstract
Nucleation of microtubules is central to assembly of the mitotic spindle, which is required for each cell division. gamma-Tubulin is a universal component essential for microtubule nucleation from centrosomes. To elucidate the mechanism of microtubule nucleation in budding yeast we reconstituted and characterized the yeast gamma-tubulin complex (Tub4p complex) produced in insect cells. The recombinant complex has the same sedimentation coefficient (11.6 S) as the native complex in yeast cell extracts and contains one molecule of Spc97p, one molecule of Spc98p, and two molecules of Tub4p. The reconstituted Tub4p complex binds preformed microtubules and has a low nucleating activity, allowing us to begin a detailed analysis of conditions that enhance this nucleating activity. We tested whether binding of the recombinant Tub4p complex to the spindle pole body docking protein Spc110p affects its nucleating activity. The solubility of recombinant Spc110p in insect cells is improved by coexpression with yeast calmodulin (Cmd1p). The Spc110p/Cmd1p complex has a small sedimentation coefficient (4.2 S) and a large Stokes radius (14.3 nm), indicative of an elongated structure. The Tub4p complex binds Spc110p/Cmd1p via Spc98p and the K(d) for binding is 150 nM. The low nucleation activity of the Tub4p complex is not enhanced when it is bound to Spc110p/Cmd1p, suggesting that it requires additional components or modifications to achieve robust activity. Finally, we report the identification of a large 22 S Tub4p complex in yeast extract that contains multimers of Spc97p similar to gamma-tubulin ring complexes found in higher eukaryotic cells.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.