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体外和体内DNA环化的定量比较:染色质在短距离内增加了有效的DNA柔韧性。

Quantitative comparison of DNA looping in vitro and in vivo: chromatin increases effective DNA flexibility at short distances.

作者信息

Ringrose L, Chabanis S, Angrand P O, Woodroofe C, Stewart A F

机构信息

Gene Expression Program, European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69117 Heidelberg, Germany.

出版信息

EMBO J. 1999 Dec 1;18(23):6630-41. doi: 10.1093/emboj/18.23.6630.

Abstract

The probability that two sites on a linear DNA molecule will contact each other by looping depends on DNA flexibility. Although the flexibility of naked DNA in vitro is well characterized, looping in chromatin is poorly understood. By extending existing theory, we present a single equation that describes DNA looping over all distances. We also show that DNA looping in vitro can be measured accurately by FLP recombination between sites from 74 bp to 15 kb apart. In agreement with previous work, a persistence length of 50 nm was determined. FLP recombination of the same substrates in mammalian cells showed that chromatin increases the flexibility of DNA at short distances, giving an apparent persistence length of 27 nm.

摘要

线性DNA分子上的两个位点通过成环相互接触的概率取决于DNA的柔韧性。虽然体外裸DNA的柔韧性已得到充分表征,但染色质中的成环现象却知之甚少。通过扩展现有理论,我们提出了一个描述所有距离上DNA成环的单一方程。我们还表明,通过相距74 bp至15 kb的位点之间的FLP重组,可以准确测量体外DNA成环。与之前的工作一致,确定了50 nm的持久长度。在哺乳动物细胞中对相同底物进行FLP重组表明,染色质在短距离内增加了DNA的柔韧性,表观持久长度为27 nm。

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