Histone‐ and DNA sequence‐dependent stability of nucleosomes studied by single‐pair FRET
Histone‐ and DNA sequence‐dependent stability of nucleosomes studied by single‐pair FRET
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DOI：
10.1002/cyto.a.22320
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发表时间：
2013-09
期刊：
影响因子：
3.7
通讯作者：
K. Toth;V. Boehm;Carolin Sellmann;M. Danner;Janina Hanne;M. Berg;Ina Barz;A. Gansen;J. Langowski
中科院分区：
文献类型：
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作者：
K. Toth;V. Boehm;Carolin Sellmann;M. Danner;Janina Hanne;M. Berg;Ina Barz;A. Gansen;J. Langowski
核小体结构的打开对于进入基因组DNA是必不可少的。为了研究这一过程的机制，我们通过单分子Förster共振能量转移（FRET）监测单核细胞体上各种荧光标记位置之间的距离。在这里，我们比较重组小鼠，非洲爪蟾，酵母组蛋白的核小体重建。作为DNA序列，我们比较了5S rDNA、MMTV-B序列和Widom 601 DNA的作用。通过在开口过渡中点处的盐浓度测量的稳定性对于酵母是最低的，其次是非洲爪蟾和小鼠。601 DNA序列构建更稳定的核小体，并且FRET效率的分布比在5S rDNA或MMTV-B序列上重构的那些窄。通过一个中间状态，非洲爪蟾组蛋白中发现的开放途径，可以验证的小鼠和酵母系统和不同的DNA序列，这表明一个一般的机制访问核小体DNA。© 2013国际细胞计数促进学会
Opening of the nucleosome structure is essential for accessing genomic DNA. To study the mechanism of this process, we monitor the distance between various fluorescently labeled positions on mononucleosomes by single‐molecule Förster resonance energy transfer (FRET). Here, we compare nucleosomes reconstituted from recombinant mouse, Xenopus, and yeast histones. As DNA sequences we compared, the effect of 5S rDNA, MMTV‐B sequence, and Widom 601 DNA. The stability, as measured by the salt concentration at the opening transition midpoint, is lowest for yeast, followed by Xenopus and mouse. The 601 DNA sequence builds much more stable nucleosomes and the distribution of FRET efficiencies is narrower than for those reconstituted on 5S rDNA or MMTV‐B sequences. The opening pathway through an intermediate state, as found for Xenopus histones, could be verified for the mouse and yeast systems and for the different DNA sequences, suggesting a general mechanism for accessing nucleosomal DNA. © 2013 International Society for Advancement of Cytometry