HIGHLY CONSERVED REPETITIVE DNA-SEQUENCES ARE PRESENT AT HUMAN CENTROMERES
HIGHLY CONSERVED REPETITIVE DNA-SEQUENCES ARE PRESENT AT HUMAN CENTROMERES
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DOI：
10.1073/pnas.89.5.1695
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发表时间：
1992-03-01
影响因子：
11.1
通讯作者：
MOYZIS, RK
中科院分区：
文献类型：
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作者：
GRADY, DL;RATLIFF, RL;MOYZIS, RK
通过与啮齿类动物重复DNA的高强度杂交，从人重组重复DNA文库中分离出高度保守的重复DNA序列克隆，主要由（GGAAT）n个重复序列组成。该序列是人类卫星Il和III的主要重复序列，与酿酒酵母着丝粒的核心DNA -着丝粒DNA元件（CDE） III相似。对人类末期和果蝇多丝染色体的原位杂交显示（GGAAT）n序列定位于着丝粒区。高染性研究表明（GGAAT）n序列表现出不同寻常的氢键性质。富含嘌呤的单链与双链具有相同的热稳定性。合成DNA变异的高染色性研究表明，所有含有AATGN的序列都表现出这种不寻常的热稳定性。dna迁移分析表明，特定的hela细胞核蛋白以相对亲和力bbb10识别该序列(5)。该DNA序列的极端进化保守性、着丝粒位置、不寻常的氢键特性、对特定核蛋白的高亲和力以及与从酵母中分离的功能性着丝粒的相似性表明，该序列可能是功能性人类着丝粒的一个组成部分。
Highly conserved repetitive DNA sequence clones, largely consisting of (GGAAT)n repeats, have been isolated from a human recombinant repetitive DNA library by high-stringency hybridization with rodent repetitive DNA. This sequence, the predominant repetitive sequence in human satellites Il and III, is similar to the essential core DNA of the Saccharomyces cerevisiae centromere, centromere DNA element (CDE) III. In situ hybridization to human telophase and Drosophila polytene chromosomes shows localization of the (GGAAT)n sequence to centromeric regions. Hyperchromicity studies indicate that the (GGAAT)n sequence exhibits unusual hydrogen bonding properties. The purine-rich strand alone has the same thermal stability as the duplex. Hyperchromicity studies of synthetic DNA variants indicate that all sequences with the composition (AATGN)n exhibit this unusual thermal stability. DNA-mobility-shift assays indicate that specific HeLa-cell nuclear proteins recognize this sequence with a relative affinity > 10(5). The extreme evolutionary conservation of this DNA sequence, its centromeric location, its unusual hydrogen bonding properties, its high affinity for specific nuclear proteins, and its similarity to functional centromeres isolated from yeast suggest that this sequence may be a component of the functional human centromere.