Characterization and Reconstitution of Drosophila γ-Tubulin Ring Complex Subunits
| Autor: | Yixian Zheng, Ruwanthi N. Gunawardane, Ona C. Martin, Lijun Zhang, Kimberly Dej, Akihiro Iwamatsu, Kan Cao |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2000 |
| Předmět: |
Embryo
Nonmammalian Protein subunit Amino Acid Motifs Molecular Sequence Data Fluorescent Antibody Technique Microtubules Models Biological Chromosomes Microtubule Antibody Specificity Tubulin Animals Drosophila Proteins grip Amino Acid Sequence Cloning Molecular Protein Structure Quaternary Peptide sequence Conserved Sequence In Situ Hybridization Microtubule nucleation Centrosome Tubulin complex biology Cell Biology Molecular biology Precipitin Tests Cell biology Protein Subunits Drosophila melanogaster biology.protein Insect Proteins Original Article Drosophila Microtubule-Associated Proteins Sequence Alignment Drosophila Protein γ-tubulin microtubule Protein Binding |
| Zdroj: | The Journal of Cell Biology |
| ISSN: | 1540-8140 0021-9525 |
| Popis: | The gamma-tubulin ring complex (gammaTuRC) is important for microtubule nucleation from the centrosome. In addition to gamma-tubulin, the Drosophila gammaTuRC 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 gamma-tubulin complex (gammaTuSC) and the gammaTuRC, while the remaining subunits are found only in the gammaTuRC. To study gammaTuRC assembly and function, we first reconstituted gammaTuSC using the baculovirus expression system. Using the reconstituted gammaTuSC, we showed for the first time that this subcomplex of the gammaTuRC has microtubule binding and capping activities. Next, we characterized two new gammaTuRC subunits, Dgrips128 and 163, and showed that they are centrosomal proteins. Sequence comparisons among all known gammaTuRC subunits revealed two novel sequence motifs, which we named grip motifs 1 and 2. We found that Dgrips128 and 163 can each interact with gammaTuSC. However, this interaction is insufficient for gammaTuRC assembly. |
| Databáze: | OpenAIRE |
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