Laboratory of Structural Biology, Graduate School of Advanced Science and Engineering/RISE, Waseda University, 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo 162-8640, Japan, Program in Chemistry and Life Science, School of Science and Engineering, Department of Chemistry, Graduate School of Science and Engineering, Meisei University, 2-1-1 Hodokubo, Hino, Tokyo 191-8506, Japan and Graduate School of Medical Life Science, Yokohama City University, 1-7-29 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan.
Nucleic Acids Res. 2014 Feb;42(3):1541-52. doi: 10.1093/nar/gkt1006. Epub 2013 Oct 29.
Telomeric DNAs consist of tandem repeats of G-clusters such as TTAGGG and TG1-3, which are the human and yeast repeat sequences, respectively. In the yeast Saccharomyces cerevisiae, the telomeric repeats are non-nucleosomal, whereas in humans, they are organized in tightly packaged nucleosomes. However, previous in vitro studies revealed that the binding affinities of human and yeast telomeric repeat sequences to histone octamers in vitro were similar, which is apparently inconsistent with the differences in the human and yeast telomeric chromatin structures. To further investigate the relationship between telomeric sequences and chromatin structure, we examined the effect of telomeric repeats on the formation of positioned nucleosomes in vivo by indirect end-label mapping, primer extension mapping and nucleosome repeat analyses, using a defined minichromosome in yeast cells. We found that the human and yeast telomeric repeat sequences both disfavour nucleosome assembly and alter nucleosome positioning in the yeast minichromosome. We further demonstrated that the G-clusters in the telomeric repeats are required for the nucleosome-disfavouring properties. Thus, our results suggest that this inherent structural feature of the telomeric repeat sequences is involved in the functional dynamics of the telomeric chromatin structure.
端粒 DNA 由 G-簇串联重复组成,如 TTAGGG 和 TG1-3,分别是人类和酵母的重复序列。在酵母酿酒酵母中,端粒重复是非核小体的,而在人类中,它们被组织成紧密包装的核小体。然而,以前的体外研究表明,人类和酵母端粒重复序列与体外组蛋白八聚体的结合亲和力相似,这显然与人类和酵母端粒染色质结构的差异不一致。为了进一步研究端粒序列与染色质结构之间的关系,我们通过间接末端标记作图、引物延伸作图和核小体重复分析,使用酵母细胞中的定义性小染色体,研究了端粒重复对体内定位核小体形成的影响。我们发现,人类和酵母端粒重复序列都不利于核小体组装,并改变酵母小染色体中的核小体定位。我们进一步证明,端粒重复中的 G-簇对于核小体不利的特性是必需的。因此,我们的结果表明,端粒重复序列的这种固有结构特征参与了端粒染色质结构的功能动力学。