Vertebrate kinetochore protein architecture: protein copy number.
Vertebrate kinetochore protein architecture: protein copy number.
复制标题
DOI：
10.1083/jcb.200912022
复制
发表时间：
2010-06-14
期刊：
影响因子：
--
通讯作者：
Salmon ED
中科院分区：
文献类型：
--
作者：
Johnston K;Joglekar A;Hori T;Suzuki A;Fukagawa T;Salmon ED
动粒组分的化学计量确定，表明多个微管结合脊椎动物和单个微管结合酵母动粒之间的保护。为了确定脊椎动物细胞中动粒的分子结构，我们测量了连接动粒微管（MT [kMT]）和着丝粒DNA的8种动粒蛋白的拷贝数。我们使用了荧光比率法和鸡DT 40细胞系，其中编码所分析蛋白的内源基因座被删除，并使用整合的绿色荧光蛋白融合转基因进行补充。对于中期平均4.3 kMT，对于MT结合KNL-1/Mis 12复合物/Ndc 80复合物网络的成员，每个kMT的蛋白质拷贝数在7和9之间。组成型着丝粒相关网络的四个成员：着丝粒蛋白C（CENP-C），CENP-H，CENP-I和CENP-T的基因频率在6到9之间。所有这些蛋白质的每kMT的拷贝数的相似性表明，每个MT末端连接到DNA的脊椎动物细胞中的6至9个纤维单位附着模块，一个结论，表明多个MT结合脊椎动物和单一的MT结合芽殖酵母动粒之间的建筑保护。
The stoichiometry of kinetochore components is determined, suggesting conservation between multiple microtubule-binding vertebrate and single microtubule-binding yeast kinetochores. To define the molecular architecture of the kinetochore in vertebrate cells, we measured the copy number of eight kinetochore proteins that link kinetochore microtubules (MTs [kMTs]) to centromeric DNA. We used a fluorescence ratio method and chicken DT40 cell lines in which endogenous loci encoding the analyzed proteins were deleted and complemented using integrated green fluorescent protein fusion transgenes. For a mean of 4.3 kMTs at metaphase, the protein copy number per kMT is between seven and nine for members of the MT-binding KNL-1/Mis12 complex/Ndc80 complex network. It was between six and nine for four members of the constitutive centromere-associated network: centromere protein C (CENP-C), CENP-H, CENP-I, and CENP-T. The similarity in copy number per kMT for all of these proteins suggests that each MT end is linked to DNA by six to nine fibrous unit attachment modules in vertebrate cells, a conclusion that indicates architectural conservation between multiple MT-binding vertebrate and single MT-binding budding yeast kinetochores.
登录
查看更多内容
影响因子：
3.4
作者：
Patterson, GH;Knobel, SM;Piston, DW
通讯作者：
Piston, DW
影响因子：
11.4
作者：
Fukagawa, T;Mikami, Y;Ikemura, T
通讯作者：
Ikemura, T
影响因子：
64.5
作者：
Hori, Tetsuya;Amano, Miho;Fukagawa, Tatsuo
通讯作者：
Fukagawa, Tatsuo
DOI：
10.1016/j.cub.2009.02.056
发表时间：
2009-04-28
期刊：
Current biology : CB
影响因子：
--
作者：
Joglekar AP;Bloom K;Salmon ED
通讯作者：
Salmon ED
影响因子：
4
作者：
Hori, T;Haraguchi, T;Fukagawa, T
通讯作者：
Fukagawa, T