TUB4 COMPLEX IN YEAST

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 酵母纺锤体极体（SPB）在功能上相当于高等真核细胞中的中心体。SPB是一个多层的细胞器，组织细胞核和细胞质微管。 我们正在努力重建SPB的微管成核活性。 小γ-微管蛋白复合物的特征在于包含三种蛋白质：Tub 4p，Spc 97 p和Spc 98 p。我们已经在杆状病毒中表达了所有三种蛋白质，并发现它们容易组装成稳定的可溶性复合物。然而，小复合物的成核能力相当低，这表明我们缺少成核所需的组分。酵母纺锤体极体（SPB）在功能上相当于高等真核细胞中的中心体。SPB是一个多层的细胞器，组织细胞核和细胞质微管。 我们正在努力重建SPB的微管成核活性。 小γ-微管蛋白复合物的特征在于包含三种蛋白质：Tub 4p，Spc 97 p和Spc 98 p。我们已经在杆状病毒中表达了所有三种蛋白质，并发现它们容易组装成稳定的可溶性复合物。然而，小络合物的成核能力相当低。 最近，Agard实验室（与我们合作）表明，小复合物中的Tub 4p分子相距太远，无法使微管成核。

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.