The Effect of Basepair Mismatch on DNA Strand Displacement

被引:30
作者
Broadwater, D. W. Bo, Jr. [1 ]
Kim, Harold D. [1 ]
机构
[1] Georgia Inst Technol, Sch Phys, Atlanta, GA 30332 USA
基金
美国国家卫生研究院; 美国国家科学基金会;
关键词
SINGLE-MOLECULE FLUORESCENCE; ORIGAMI MECHANISMS; ENERGY LANDSCAPE; MEAN; 1ST-PASSAGE; RANDOM-WALKS; HYBRIDIZATION; KINETICS; TIME; COMPUTATION; DESIGN;
D O I
10.1016/j.bpj.2016.02.027
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
DNA strand displacement is a key reaction in DNA homologous recombination and DNA mismatch repair and is also heavily utilized in DNA-based computation and locomotion. Despite its ubiquity in science and engineering, sequence dependent effects of displacement kinetics have not been extensively characterized. Here, we measured toehold-mediated strand displacement kinetics using single-molecule fluorescence in the presence of a single basepair mismatch. The apparent displacement rate varied significantly when the mismatch was introduced in the invading DNA strand. The rate generally decreased as the mismatch in the invader was encountered earlier in displacement. Our data indicate that a single base pair mismatch in the invader stalls branch migration and displacement occurs via direct dissociation of the destabilized incumbent strand from the substrate strand. We combined both branch migration and direct dissociation into a model, which we term the concurrent displacement model, and used the first passage time approach to quantitatively explain the salient features of the observed relationship. We also introduce the concept of splitting probabilities to justify that the concurrent model can be simplified into a three-step sequential model in the presence of an invader mismatch. We expect our model to become a powerful tool to design DNA-based reaction schemes with broad functionality.
引用
收藏
页码:1476 / 1484
页数:9
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