RAPID RENATURATION OF COMPLEMENTARY-DNA STRANDS MEDIATED BY CATIONIC DETERGENTS - A ROLE FOR HIGH-PROBABILITY BINDING DOMAINS IN ENHANCING THE KINETICS OF MOLECULAR ASSEMBLY PROCESSES
RAPID RENATURATION OF COMPLEMENTARY-DNA STRANDS MEDIATED BY CATIONIC DETERGENTS - A ROLE FOR HIGH-PROBABILITY BINDING DOMAINS IN ENHANCING THE KINETICS OF MOLECULAR ASSEMBLY PROCESSES
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DOI：
10.1073/pnas.88.18.8237
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发表时间：
1991-09-01
影响因子：
11.1
通讯作者：
BERG, P
中科院分区：
文献类型：
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作者：
PONTIUS, BW;BERG, P
通过添加简单的阳离子去污剂，互补DNA链的复性速度可以提高10(4)倍，而且这个反应在定性和定量上都与纯化的异质核糖核蛋白A1蛋白非常相似。在最佳条件下，复性速度比在68℃的1M氯化钠中进行的反应快2000倍。该反应是关于DNA浓度的二级反应，反应速率接近或等于互补链在溶液中彼此相遇的速率。复性甚至可以观察到远高于预期的双链DNA熔融温度，表明一些阳离子洗涤剂具有DNA双螺旋稳定特性。在存在高达10(6)倍的非互补序列时，反应也非常迅速，证实复性是特异的，并且相对独立于异源DNA。这一发现还意味着，每条链每秒可以采样多达数千个潜在的靶序列。这类试剂可能对需要快速核酸复性的程序有用，这些结果表明可以识别和设计其他提高缔合反应动力学的化合物。此外，这项工作进一步支持了一个模型，该模型将柔性的、弱相互作用的、重复的结构域的存在与它们在体外和体内快速分子组装过程中的功能联系起来。
The rate of renaturation for complementary DNA strands can be enhanced > 10(4)-fold by the addition of simple cationic detergents, and the reaction is qualitatively and quantitatively very similar to that found with purified heterogeneous nuclear ribonucleoprotein A1 protein. Under optimal conditions, renaturation rates are > 2000-fold faster than reactions run in 1 M NaCl at 68-degrees-C. The reaction is second-order with respect to DNA concentration, and reaction rates approach or equal the rate with which complementary strands are expected to encounter each other in solution. Renaturation can even be observed well above the expected melting temperature of the duplex DNA, demonstrating that some cationic detergents have DNA double-helix-stabilizing properties. The reaction is also extremely rapid in the presence of up to a 10(6)-fold excess of noncomplementary sequences, establishing that renaturation is specific and relatively independent of heterologous DNA. This finding also implies that up to several thousand potential target sequences can be sampled per strand per second. Such reagents may be useful for procedures that require rapid nucleic acid renaturation, and these results suggest ways to identify and design other compounds that increase the kinetics of association reactions. Moreover, this work provides further support for a model relating the existence of flexible, weakly interacting, repeating domains to their function in rapid molecular assembly processes in vitro and in vivo.