TUB4 COMPLEX IN YEAST

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 酵母纺锤体极体(SPB)在功能上等同于高等真核细胞中的中心体。SPB是一个多层细胞器，既有核的也有细胞质的微管。我们正在努力重建SPB的微管成核活性。鉴定良好的小分子微管蛋白复合体包含三种蛋白质：Tub4p、Spc97p和Spc98p。我们已经在杆状病毒中表达了这三种蛋白，并发现它们很容易组装成稳定的、可溶的复合体。然而，小复合体的成核能力相当低，这表明我们缺少成核所需的成分。酵母纺锤体极体(SPB)在功能上等同于高等真核细胞中的中心体。SPB是一个多层细胞器，既有核的也有细胞质的微管。我们正在努力重建SPB的微管成核活性。鉴定良好的小分子微管蛋白复合体包含三种蛋白质：Tub4p、Spc97p和Spc98p。我们已经在杆状病毒中表达了这三种蛋白，并发现它们很容易组装成稳定的、可溶的复合体。然而，小分子络合物的成核能力很低。最近，阿加德实验室(与我们合作)显示，小复合体中的Tub4p分子相距太远，无法形成微管。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The yeast spindle pole body (SPB) is functionally equivalent to the centrosome in higher eukaryotic cells. The SPB is a multilayered organelle that organizes both the nuclear and cytoplasmic microtubules. We are working toward reconstitution of the microtubule nucleating activity of the SPB. The well characterized small gamma-tubulin complex contains three proteins: Tub4p, Spc97p, and Spc98p. We have expressed all three proteins in baculovirus and found that they readily assemble into a stable, soluble complex. However, the nucleation capacity of the small complex is quite low, suggesting we are missing components required for nucleation. The yeast spindle pole body (SPB) is functionally equivalent to the centrosome in higher eukaryotic cells. The SPB is a multilayered organelle that organizes both the nuclear and cytoplasmic microtubules. We are working toward reconstitution of the microtubule nucleating activity of the SPB. The well characterized small gamma-tubulin complex contains three proteins: Tub4p, Spc97p, and Spc98p. We have expressed all three proteins in baculovirus and found that they readily assemble into a stable, soluble complex. However, the nucleation capacity of the small complex is quite low. Recently the Agard lab (in collaboration with us) showed that the Tub4p molecules in the small complex are too far apart to nucleate microtubules.