Khan W A, Chisholm R, Tadayyon S, Subasinghe A, Norton P, Samarabandu J, Johnston L J, Knoll J H, Rogan P K
Department of Pathology, Schulich School of Medicine and Dentistry, London, Ont, Canada.
Cytogenet Genome Res. 2013;139(4):234-42. doi: 10.1159/000348744. Epub 2013 Mar 23.
Despite extensive analyses on the centromere and its associated proteins, detailed studies of centromeric DNA structure have provided limited information about its topography in condensed chromatin. We have developed a method with correlative fluorescence light microscopy and atomic force microscopy that investigates the physical and structural organization of α-satellite DNA sequences in the context of its associated protein, CENP-B, on human metaphase chromosome topography. Comparison of centromeric DNA and protein distribution patterns in fixed homologous chromosomes indicates that CENP-B and α-satellite DNA are distributed distinctly from one another and relative to observed centromeric ridge topography. Our approach facilitates correlated studies of multiple chromatin components comprising higher-order structures of human metaphase chromosomes.
尽管对着丝粒及其相关蛋白进行了广泛分析，但对着丝粒DNA结构的详细研究提供的关于其在浓缩染色质中的拓扑结构的信息有限。我们开发了一种结合荧光显微镜和原子力显微镜的方法，该方法在人类中期染色体拓扑结构的背景下，研究了α卫星DNA序列与其相关蛋白CENP - B的物理和结构组织。固定的同源染色体中着丝粒DNA和蛋白质分布模式的比较表明，CENP - B和α卫星DNA彼此之间以及相对于观察到的着丝粒脊拓扑结构的分布是不同的。我们的方法有助于对构成人类中期染色体高阶结构的多种染色质成分进行相关研究。
