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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...
Autores principales: | , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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The Rockefeller University Press
2000
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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. |
format | Text |
id | pubmed-2150673 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2000 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
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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