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Characterization and Reconstitution of Drosophila γ-Tubulin Ring Complex Subunits

The γ-tubulin ring complex (γTuRC) is important for microtubule nucleation from the centrosome. In addition to γ-tubulin, the Drosophila γTuRC contains at least six subunits, three of which [Drosophila gamma ring proteins (Dgrips) 75/d75p, 84, and 91] have been characterized previously. Dgrips84 and...

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Autores principales: Gunawardane, Ruwanthi N., Martin, Ona C., Cao, Kan, Zhang, Lijun, Dej, Kimberly, Iwamatsu, Akihiro, Zheng, Yixian
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2000
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2150673/
https://www.ncbi.nlm.nih.gov/pubmed/11134079
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author Gunawardane, Ruwanthi N.
Martin, Ona C.
Cao, Kan
Zhang, Lijun
Dej, Kimberly
Iwamatsu, Akihiro
Zheng, Yixian
author_facet Gunawardane, Ruwanthi N.
Martin, Ona C.
Cao, Kan
Zhang, Lijun
Dej, Kimberly
Iwamatsu, Akihiro
Zheng, Yixian
author_sort Gunawardane, Ruwanthi N.
collection PubMed
description The γ-tubulin ring complex (γTuRC) is important for microtubule nucleation from the centrosome. In addition to γ-tubulin, the Drosophila γTuRC contains at least six subunits, three of which [Drosophila gamma ring proteins (Dgrips) 75/d75p, 84, and 91] have been characterized previously. Dgrips84 and 91 are present in both the small γ-tubulin complex (γTuSC) and the γTuRC, while the remaining subunits are found only in the γTuRC. To study γTuRC assembly and function, we first reconstituted γTuSC using the baculovirus expression system. Using the reconstituted γTuSC, we showed for the first time that this subcomplex of the γTuRC has microtubule binding and capping activities. Next, we characterized two new γTuRC subunits, Dgrips128 and 163, and showed that they are centrosomal proteins. Sequence comparisons among all known γTuRC subunits revealed two novel sequence motifs, which we named grip motifs 1 and 2. We found that Dgrips128 and 163 can each interact with γTuSC. However, this interaction is insufficient for γTuRC assembly.
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spelling pubmed-21506732008-05-01 Characterization and Reconstitution of Drosophila γ-Tubulin Ring Complex Subunits Gunawardane, Ruwanthi N. Martin, Ona C. Cao, Kan Zhang, Lijun Dej, Kimberly Iwamatsu, Akihiro Zheng, Yixian J Cell Biol Original Article The γ-tubulin ring complex (γTuRC) is important for microtubule nucleation from the centrosome. In addition to γ-tubulin, the Drosophila γTuRC contains at least six subunits, three of which [Drosophila gamma ring proteins (Dgrips) 75/d75p, 84, and 91] have been characterized previously. Dgrips84 and 91 are present in both the small γ-tubulin complex (γTuSC) and the γTuRC, while the remaining subunits are found only in the γTuRC. To study γTuRC assembly and function, we first reconstituted γTuSC using the baculovirus expression system. Using the reconstituted γTuSC, we showed for the first time that this subcomplex of the γTuRC has microtubule binding and capping activities. Next, we characterized two new γTuRC subunits, Dgrips128 and 163, and showed that they are centrosomal proteins. Sequence comparisons among all known γTuRC subunits revealed two novel sequence motifs, which we named grip motifs 1 and 2. We found that Dgrips128 and 163 can each interact with γTuSC. However, this interaction is insufficient for γTuRC assembly. The Rockefeller University Press 2000-12-25 /pmc/articles/PMC2150673/ /pubmed/11134079 Text en © 2000 The Rockefeller University Press This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Original Article
Gunawardane, Ruwanthi N.
Martin, Ona C.
Cao, Kan
Zhang, Lijun
Dej, Kimberly
Iwamatsu, Akihiro
Zheng, Yixian
Characterization and Reconstitution of Drosophila γ-Tubulin Ring Complex Subunits
title Characterization and Reconstitution of Drosophila γ-Tubulin Ring Complex Subunits
title_full Characterization and Reconstitution of Drosophila γ-Tubulin Ring Complex Subunits
title_fullStr Characterization and Reconstitution of Drosophila γ-Tubulin Ring Complex Subunits
title_full_unstemmed Characterization and Reconstitution of Drosophila γ-Tubulin Ring Complex Subunits
title_short Characterization and Reconstitution of Drosophila γ-Tubulin Ring Complex Subunits
title_sort characterization and reconstitution of drosophila γ-tubulin ring complex subunits
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2150673/
https://www.ncbi.nlm.nih.gov/pubmed/11134079
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