Souès S, Adams I R
Laboratory of Molecular Biology, Medical Research Council Centre, Hills Road, Cambridge CB2 2QH, UK.
J Cell Sci. 1998 Sep;111 ( Pt 18):2809-18. doi: 10.1242/jcs.111.18.2809.
The monoclonal antibody 78H6 recognises an 85 kDa component of the yeast spindle pole body. Here we identify and characterise this component as Spc72p, the product of YAL047C. The sequence of SPC72 contains potential coiled-coil domains; its overexpression induced formation of large polymers that were strictly localised at the outer plaque and at the bridge of the spindle pole body. Immunoelectron microscopy confirmed that Spc72p was a component of these polymers. SPC72 was found to be non-essential for cell growth, but its deletion resulted in abnormal spindle positioning, aberrant nuclear migration and defective mating capability. Precisely, deletion of SPC72 resulted in a decreased number of astral microtubules: early in the cell cycle only few were detectable, and these were unattached to the spindle pole body in small-budded cells. Later in the cell cycle few, if any, remained, and they were unable to align the spindle properly. We conclude that Spc72p is not absolutely required for nucleation per se, but is needed for normal abundance and stability of astral microtubules.
单克隆抗体78H6可识别酵母纺锤体极体的一个85 kDa的组分。在此,我们鉴定并表征该组分为Spc72p,即YAL047C的产物。SPC72的序列包含潜在的卷曲螺旋结构域;其过表达诱导形成大聚合物,这些聚合物严格定位于纺锤体极体的外斑和桥处。免疫电子显微镜证实Spc72p是这些聚合物的一个组分。发现SPC72对细胞生长并非必需,但缺失它会导致纺锤体定位异常、核迁移异常以及交配能力缺陷。确切地说,SPC72的缺失导致星体微管数量减少:在细胞周期早期仅能检测到少数星体微管,并且在小芽殖细胞中这些微管未附着于纺锤体极体。在细胞周期后期,几乎没有星体微管留存,并且它们无法正确排列纺锤体。我们得出结论,Spc72p本身对于成核并非绝对必需,但对于星体微管的正常丰度和稳定性是必需的。