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植物亚端粒处两种异染色质形式之间的转变。

Transition between two forms of heterochromatin at plant subtelomeres.

作者信息

Sýkorová E, Fajkus J, Ito M, Fukui K

机构信息

Institute of Biophysics, Czech Academy of Sciences and Department of Analysis of Biologically Important Molecular Complexes, Masaryk University Brno, Czech Republic.

出版信息

Chromosome Res. 2001;9(4):309-23. doi: 10.1023/a:1016698713959.

Abstract

The manner of packing of the terminal DNA loci into nucleosomes and higher order structures may strongly influence their functional interactions. Besides the structural flexibility of telomeric DNA sequences, conserved features of their chromatin including short nucleosome phasing (157 bp) and nucleosome sliding have been described previously. To gain a complementary knowledge of subtelomeres, we have analysed the chromatin structure of two subtelomeric tandem repeats from the plant Silene latifolia: X43.1 and 15Ssp. X43.1 shows two distinct nucleosome periodicities--157 and 188 bp. Preferred positions of its two nucleosomes have been mapped at both low and high resolution and the experimental results correspond to computer-predicted positions. 15Ssp is a newly-discovered sequence showing a telomere-associated position by PCR and a subtelomeric location by pulsed-field gel electrophoresis and fluorescence in situ hybridisation. Its 159 bp sequence unit shows a tandem arrangement and the presence of micrococcal nuclease-hypersensitive sites when either naked DNA or chromatin is digested. Use of a chemical nuclease results in a regular nucleosome ladder of 157 bp periodicity. Moreover, 15Ssp mononucleosomes show instability and absence of specific positioning, features typical for telomeric chromatin.

摘要

端粒DNA位点包装成核小体和更高层次结构的方式可能会强烈影响它们的功能相互作用。除了端粒DNA序列的结构灵活性外,其染色质的保守特征,包括短核小体相位(157 bp)和核小体滑动,此前已有描述。为了获得亚端粒的补充知识,我们分析了来自植物宽叶蝇子草的两个亚端粒串联重复序列的染色质结构:X43.1和15Ssp。X43.1显示出两种不同的核小体周期——157和188 bp。已在低分辨率和高分辨率下绘制了其两个核小体的优选位置,实验结果与计算机预测位置相符。15Ssp是一个新发现的序列,通过PCR显示与端粒相关的位置,通过脉冲场凝胶电泳和荧光原位杂交显示亚端粒位置。其159 bp序列单元呈串联排列,当消化裸露DNA或染色质时存在微球菌核酸酶超敏位点。使用化学核酸酶会产生157 bp周期的规则核小体梯带。此外,15Ssp单核小体表现出不稳定性且缺乏特定定位,这是端粒染色质的典型特征。

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