Keates R A, Sarangi F, Ling V
Proc Natl Acad Sci U S A. 1981 Sep;78(9):5638-42. doi: 10.1073/pnas.78.9.5638.
We have examined mutant lines of Chinese hamster ovary cells that have increased resistance to the antimicrotubule drug Colcemid. Analysis of the functional properties of purified microtubule protein indicates that increased tolerance to the drug in vivo is reflected in altered properties of microtubules and tubulin in vitro. In this study, we have examined one series of related mutants and have found different microtubule alterations associated with each selection step. These changes include decreased Colcemid-binding affinity, an altered electrophoretic pattern of tubulin subcomponents, increased resistance to Colcemid inhibition of polymerization in vitro and, in one case, a decreased critical concentration for microtubule assembly. Characterized mutants of the class described here will be useful for probing the regulation of microtubule assembly in vivo.
我们已经检测了对抗微管药物秋水仙酰胺具有增强抗性的中国仓鼠卵巢细胞突变株系。对纯化的微管蛋白功能特性的分析表明,体内对该药物耐受性的增加反映在体外微管和微管蛋白特性的改变上。在本研究中,我们检测了一系列相关突变体,并发现每个选择步骤都伴随着不同的微管变化。这些变化包括秋水仙酰胺结合亲和力降低、微管蛋白亚组分电泳图谱改变、体外对秋水仙酰胺抑制聚合作用的抗性增加,以及在一个案例中微管组装临界浓度降低。这里描述的这类经过表征的突变体将有助于探究体内微管组装的调控机制。