The constitutive centromere component CENP-50 is required for recovery from spindle damage
The constitutive centromere component CENP-50 is required for recovery from spindle damage
复制标题
DOI：
10.1128/mcb.25.23.10315-10328.2005
复制
发表时间：
2005-12-01
影响因子：
5.3
通讯作者：
Fukagawa, T
中科院分区：
文献类型：
--
作者：
Minoshima, Y;Hori, T;Fukagawa, T
我们将CENP-50鉴定为一种新的动粒组分。我们发现CENP-50是着丝粒的组成成分，其在脊椎动物细胞的整个细胞周期中与CENP-A和CENP-H共定位。为了确定CENP-50的确切作用，我们通过在鸡DT 40细胞系中产生功能丧失突变体来检查其在着丝粒功能中的作用。CENP-50敲除不是致命的;然而，具有这种突变的细胞的生长速率比野生型细胞的生长速率慢。我们观察到CENP-50缺陷细胞完成有丝分裂的时间比野生型细胞长。CENP-50的着丝粒定位在CENP-H-和CENP-I-缺陷细胞中均被消除。免疫共沉淀实验表明，CENP-50与CENP-H/CENP-I复合物在鸡DT 40细胞中相互作用。我们还观察到CENP-50缺陷细胞中的严重有丝分裂缺陷，当有丝分裂检查点被激活时具有明显的过早姐妹染色单体分离，表明CENP-50是从纺锤体损伤中恢复所需的。
We identified CENP-50 as a novel kinetochore component. We found that CENP-50 is a constitutive component of the centromere that colocalizes with CENP-A and CENP-H throughout the cell cycle in vertebrate cells. To determine the precise role of CENP-50, we examined its role in centromere function by generating a loss-of-function mutant in the chicken DT40 cell line. The CENP-50 knockout was not lethal; however, the growth rate of cells with this mutation was slower than that of wild-type cells. We observed that the time for CENP-50-deficient cells to complete mitosis was longer than that for wild-type cells. Centromeric localization of CENP-50 was abolished in both CENP-H- and CENP-I-deficient cells. Coimmunoprecipitation experiments revealed that CENP-50 interacted with the CENP-H/CENP-I complex in chicken DT40 cells. We also observed severe mitotic defects in CENP-50-deficient cells with apparent premature sister chromatid separation when the mitotic checkpoint was activated, indicating that CENP-50 is required for recovery from spindle damage.