Densmore L, Payne W E, Fitzgerald-Hayes M
Department of Biochemistry, University of Massachusetts, Amherst 01003.
Mol Cell Biol. 1991 Jan;11(1):154-65. doi: 10.1128/mcb.11.1.154-165.1991.
We have used in vivo genomic footprinting to investigate the protein-DNA interactions within the conserved DNA elements (CDEI, CDEII, and CDEIII) in the centromere from chromosome III of the yeast Saccharomyces cerevisiae. The in vivo footprint pattern obtained from wild-type cells shows that some guanines within the centromere DNA are protected from methylation by dimethyl sulfate. These results are consistent with studies demonstrating that yeast cells contain sequence-specific centromere DNA-binding proteins. Our in vivo experiments on chromosomes with mutant centromeres show that some mutations which affect chromosome segregation also alter the footprint pattern caused by proteins bound to the centromere DNA. The results of this study provide the first fine-structure map of proteins bound to centromere DNA in living yeast cells and suggest a direct correlation between these protein-DNA interactions and centromere function.
我们利用体内基因组足迹法来研究酿酒酵母第三条染色体着丝粒中保守DNA元件（CDEI、CDEII和CDEIII）内的蛋白质-DNA相互作用。从野生型细胞获得的体内足迹模式表明，着丝粒DNA内的一些鸟嘌呤受到硫酸二甲酯的甲基化保护。这些结果与证明酵母细胞含有序列特异性着丝粒DNA结合蛋白的研究一致。我们对具有突变着丝粒的染色体进行的体内实验表明，一些影响染色体分离的突变也会改变由与着丝粒DNA结合的蛋白质引起的足迹模式。这项研究的结果提供了第一张活酵母细胞中与着丝粒DNA结合的蛋白质的精细结构图，并表明这些蛋白质-DNA相互作用与着丝粒功能之间存在直接关联。
