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用于重组酶介导的DNA联会的随机游走模型。

Random walk models for DNA synapsis by resolvase.

作者信息

Sessions R B, Oram M, Szczelkun M D, Halford S E

机构信息

Department of Biochemistry, University of Bristol, UK.

出版信息

J Mol Biol. 1997 Jul 18;270(3):413-25. doi: 10.1006/jmbi.1997.1108.

Abstract

During site-specific recombination by resolvase, the protein binds to two sites on a supercoiled DNA molecule and the loaded sites then interact with each other to form a synaptic complex. The kinetics of synapsis show non-exponential behaviour extending over five log units of time and are independent of the length of the DNA molecule and the length of DNA between the sites. In this study, numerical models were developed in order to account for how fluctuations in the structure of supercoiled DNA might lead to the juxtaposition of distant sites in a manner consistent with the experimental data on synapsis by resolvase. Models where the juxtaposition arises from fluctuations around branch points in the superhelix failed to match the data: they yielded non-exponential kinetics but only over two log units of time and they predicted longer synapsis times for both larger DNA molecules and larger inter-site spacings. In another model, one fraction of the juxtaposition events gives rise directly to the productive complex while the remaining fraction initially yields a non-productive complex: the latter molecules undergo no further fluctuations until the abortive synapse dissociates at the end of a delay period. This model again failed to match the experimental data. However, the inclusion of three sorts of non-productive complexes, each with a different delay constant, led to progress curves that concurred with the data. Schemes were also developed to account for the juxtaposition of three sites at a branch point in supercoiled DNA.

摘要

在解离酶进行位点特异性重组的过程中,该蛋白质会结合到超螺旋DNA分子上的两个位点,然后已结合位点相互作用形成一个突触复合体。突触形成的动力学表现出非指数行为,时间跨度达五个对数单位,且与DNA分子的长度以及位点间DNA的长度无关。在本研究中,开发了数值模型,以解释超螺旋DNA结构的波动如何可能导致远距离位点并列,其方式与解离酶介导突触形成的实验数据一致。超螺旋分支点周围波动导致并列的模型无法与数据匹配:它们产生的非指数动力学仅在两个对数单位的时间内出现,并且它们预测对于更大的DNA分子和更大的位点间距,突触形成时间会更长。在另一个模型中,一部分并列事件直接产生有活性的复合体,而其余部分最初产生无活性的复合体:后者分子在延迟期结束时无效突触解离之前不会进一步波动。该模型再次无法与实验数据匹配。然而,纳入三种具有不同延迟常数的无活性复合体,得到的进展曲线与数据相符。还开发了一些方案来解释超螺旋DNA分支点处三个位点的并列情况。

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