Measuring the elasticity of ribonucleotide(s)-containing DNA molecules using AFM.
Measuring the elasticity of ribonucleotide(s)-containing DNA molecules using AFM.
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DOI：
10.1007/978-1-4939-2562-9_3
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发表时间：
2015-01-01
期刊：
影响因子：
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通讯作者：
Storici, Francesca
中科院分区：
文献类型：
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作者：
Koh, Kyung Duk;Chiu, Hsiang-Chih;Storici, Francesca
核糖核苷酸，即核糖核苷单磷酸（rNMPs），可能是基因组DNA中最非规范的核苷酸。rNMPs要么在DNA复制过程中没有从冈崎片段中移除，要么被纳入并分散在整个基因组中，对DNA的结构造成扰动，并威胁到DNA的稳定性。DNA的不稳定性主要是由于rNMPs中额外的2'-羟基（OH）基团引起的局部结构效应，可能干扰细胞内的各种分子相互作用。由于rNMPs的这些结构扰动，DNA的弹性特性也受到影响。在这里，我们展示了一种方法来测试DNA双链中rNMP的存在是否会改变DNA的弹性，方法是通过实施基于原子力显微镜（AFM）的单分子力测量含有短rNMP的寡核苷酸（oligos）。
Ribonucleotides, ribonucleoside monophosphates (rNMPs), have been revealed as possibly the most noncanonical nucleotides in genomic DNA. rNMPs, either not removed from Okazaki fragments during DNA replication or incorporated and scattered throughout the genome, pose a perturbation to the structure and a threat to the stability of DNA. The instability of DNA is mainly due to the extra 2'-hydroxyl (OH) group of rNMPs which give rise to local structural effects, which may disturb various molecular interactions in cells. As a result of these structural perturbations by rNMPs, the elastic properties of DNA are also affected. Here, we show the approach to test whether the presence of rNMPs in DNA duplexes could alter the elasticity of DNA by implementing atomic force microscopy (AFM)-based single molecule force-measurements of short rNMP(s)-containing oligonucleotides (oligos).