Solubilization of Aster-Forming Proteins from Yeast: Possible Constituents of Spindle Pole Body and Reconstitution of Asters In Vitro

Cell Structure and Function

Michio Matsuhashi³  Makari Yamasaki¹  Ayumu Yamamoto³  Kazuo Nagai² 

[1] Department of Agricultural Chemistry, University of Tokyo;Department of Bioengineering, University of Tokyo Institute of Technology;Institute of Applied Microbiology, University of Tokyo

关键词: spindle pole body;    microtubules;    Saccharomyces cerevisiae;    aster;   

DOI  :  10.1247/csf.15.221

学科分类：分子生物学，细胞生物学和基因

来源: Japan Society for Cell Biology

【 摘 要 】

References(24)Cited-By(2)Spindle pole bodies (SPBs) attached to nuclei were isolated from Saccharomyces cerevisiae; they nucleated microtubules to form asters in vitro. This aster-forming activity was stable in 30%dimethylsulfoxide or 10% 2-propanol and could be solubilized with KC1. The KC1 extract contained many protein components, which aggregated upon dialysis against a low concentration salt solution. When incubated with tubulin, the aggregates formed asters. Measurements of the elongation rates of the astral microtubules indicated that the microtubules were nucleated from the SPBsor from the aggregates reconstructed from the KC1 extract by dialysis.The plus end was distal to the astral center, as in the case of the microtubule organizing center (MTOC) of mammalian cells. We suggest that the proteins extracted with KC1 are responsible for microtubule nucleation in SPBs and that this SPBseems to have the same mechanism for microtubule nucleation as the MTOC in higher eukaryotes.

【 授权许可】

Unknown