Biophysical characterization of DNA binding from single molecule force measurements.

Biophysical characterization of DNA binding from single molecule force measurements.
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DOI:
10.1016/j.plrev.2010.06.001
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发表时间:
2010-09
影响因子:
11.7
通讯作者:
Williams, Mark C.
Williams, Mark C.
中科院分区:
生物学2区
文献类型:
--
作者:
Chaurasiya, Kathy R.;Paramanathan, Thayaparan;McCauley, Micah J.;Williams, Mark C.

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单分子力谱是一种利用DNA的力学性质来研究DNA相互作用的强有力的方法。在这里,我们描述了DNA拉伸实验如何定量表征小分子和蛋白质的DNA结合。小分子表现出多种DNA结合模式,包括结合到大沟和小沟中以及插入双链DNA(dsDNA)的碱基对之间。组蛋白结合并包装dsDNA,而其他核蛋白如高迁移率族蛋白结合骨架并弯曲dsDNA。单链DNA(ssDNA)结合蛋白沿着dsDNA滑动以在复制期间定位和稳定ssDNA。其他蛋白质表现出与dsDNA和ssDNA两者的结合。核酸伴侣蛋白可以在dsDNA和ssDNA结合模式之间快速切换,而DNA聚合酶在复制叉的不同结合位点以高亲和力结合两种形式的DNA。单分子力测量定量表征这些DNA结合机制,阐明小分子相互作用和蛋白质功能。
Single molecule force spectroscopy is a powerful method that uses the mechanical properties of DNA to explore DNA interactions. Here we describe how DNA stretching experiments quantitatively characterize the DNA binding of small molecules and proteins. Small molecules exhibit diverse DNA binding modes, including binding into the major and minor grooves and intercalation between base pairs of double-stranded DNA (dsDNA). Histones bind and package dsDNA, while other nuclear proteins such as high mobility group proteins bind to the backbone and bend dsDNA. Single-stranded DNA (ssDNA) binding proteins slide along dsDNA to locate and stabilize ssDNA during replication. Other proteins exhibit binding to both dsDNA and ssDNA. Nucleic acid chaperone proteins can switch rapidly between dsDNA and ssDNA binding modes, while DNA polymerases bind both forms of DNA with high affinity at distinct binding sites at the replication fork. Single molecule force measurements quantitatively characterize these DNA binding mechanisms, elucidating small molecule interactions and protein function.
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