Ribeiro Susana Abreu, Vagnarelli Paola, Earnshaw William C
Wellcome Trust Centre for Cell Biology, University of Edinburgh, Michael Swann Building, King's Buildings, Mayfield Road, Edinburgh, EH9 3JR, Scotland, UK.
Chromosome Res. 2014 Apr;22(1):7-13. doi: 10.1007/s10577-014-9410-3.
In order to understand the three-dimensional structure of the functional kinetochore in vertebrates, we require a complete list and stoichiometry for the protein components of the kinetochore, which can be provided by genetic and proteomic experiments. We also need to know how the chromatin-containing CENP-A, which makes up the structural foundation for the kinetochore, is folded, and how much of that DNA is involved in assembling the kinetochore. In this MS, we demonstrate that functioning metaphase kinetochores in chicken DT40 cells contain roughly 50 kb of DNA, an amount that corresponds extremely closely to the length of chromosomal DNA associated with CENP-A in ChIP-seq experiments. Thus, during kinetochore assembly, CENP-A chromatin is compacted into the inner kinetochore plate without including significant amounts of flanking pericentromeric heterochromatin.
为了了解脊椎动物中功能性动粒的三维结构，我们需要动粒蛋白质成分的完整列表和化学计量，这可由遗传学和蛋白质组学实验提供。我们还需要知道构成动粒结构基础的含染色质的CENP-A是如何折叠的，以及有多少该DNA参与了动粒的组装。在本手稿中，我们证明鸡DT40细胞中发挥功能的中期动粒包含约50 kb的DNA，这一数量与ChIP-seq实验中与CENP-A相关的染色体DNA长度极为接近。因此，在动粒组装过程中，CENP-A染色质被压缩到动粒内板中，而不包括大量侧翼着丝粒周围异染色质。
