In vitro assembly of a positioned nucleosome near the hypersensitive region in simian virus 40 chromatin.
In vitro assembly of a positioned nucleosome near the hypersensitive region in simian virus 40 chromatin.
复制标题
猿猴病毒 40 染色质超敏感区域附近定位核小体的体外组装。
DOI：
10.1016/0022-2836(91)80176-u
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发表时间：
1991
影响因子：
5.6
通讯作者：
M. Bina
中科院分区：
文献类型：
--
作者：
J. H. Powers;M. Bina
以前的研究已经确定了一个核小体附近的一个潜在的晚期边界的核酸酶过敏区在猿猴病毒40染色质。我们已经performedin vitroreconstitution分析，以确定是否潜在的DNA序列编码该核小体的组装和应用羟基自由基和DNA酶I足迹技术，以检查重建的核小体的结构。这两种方法都揭示了一个精确定位的核小体在体外形成，在一个片段上跨越了以前在病毒染色质中确定的强蛋白活小体定位位点。定位的核小体的中心映射在猿病毒40 DNA上的核苷酸384和387之间。相应的核小体核心包括主要的晚期转录位点（核心内有12个碱基对）、MspI位点和先前显示的在没有蛋白质的情况下采用弯曲结构的片段。羟基自由基在掺入核小体的弯曲DNA上产生一种非常明确的切割模式。该核小体中DNA的优势周期为每圈10·26个碱基对。OH切割位点在定位核小体中的分布为以下模型提供了强有力的支持：富含A/T的片段的小沟朝向组蛋白核心，而富含G/C的序列的小沟朝外。
Previous studies have identified a nucleosome near a potential late boundary for the nuclease-hypersensitive region in simian virus 40 chromatin. We have performedin vitroreconstitution analysis to determine whether the underlying DNA sequence encodes for the assembly of this nucleosome and applied hydroxyl radical and DNase I footprinting techniques to examine the structure of the reconstituted nucleosome. Both methods revealed the formation of a precisely positioned nucleosomein vitro, on a fragment spanning the strongin vivonucleosome location site determined previously in the viral chromatin. The center of the positioned nucleosome maps between nucleotide 384 and 387 on simian virus 40 DNA. The corresponding nucleosome core includes the major-late transcription site (12 base-pairs within the core), theMspI site, and a segment shown previously to adopt a bent structure in the absence of proteins. The hydroxyl radical produces a strikingly well-defined cleavage pattern over the bent DNA incorporated in nucleosomes. The dominant periodicity of DNA in this nucleosome is 10·26 base-pairs per turn. The distribution of the ·OH cut sites in the positioned nucleosome provides strong support for models in which the minor grooves of the A/T-rich tracts are oriented toward the histone core while the minor grooves of the G/C-rich sequences are facing outward.
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DOI：
10.1073/pnas.86.19.7418
发表时间：
1989-10-01
影响因子：
11.1
作者：
SHRADER, TE;CROTHERS, DM
通讯作者：
CROTHERS, DM
影响因子：
5.6
作者：
DREW, HR;TRAVERS, AA
通讯作者：
TRAVERS, AA
DOI：
10.1073/pnas.83.15.5469
发表时间：
1986-08-01
影响因子：
11.1
作者：
TULLIUS, TD;DOMBROSKI, BA
通讯作者：
DOMBROSKI, BA
影响因子：
5.6
作者：
SATCHWELL, SC;DREW, HR;TRAVERS, AA
通讯作者：
TRAVERS, AA