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嵌入剂可促进RecA蛋白与双链DNA的结合。

Intercalators promote the binding of RecA protein to double-stranded DNA.

作者信息

Thresher R J, Griffith J D

机构信息

Lineberger Cancer Research Center, University of North Carolina Medical School, Chapel Hill 27514.

出版信息

Proc Natl Acad Sci U S A. 1990 Jul;87(13):5056-60. doi: 10.1073/pnas.87.13.5056.

DOI:10.1073/pnas.87.13.5056
PMID:2195545
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC54260/
Abstract

Ethidium bromide, acridine orange, 4'-(9-acridinylamino)methanesulfon-o-anisidide (o-AMSA), and m-AMSA induce the rapid binding of RecA protein to double-stranded (ds) DNA. The filaments formed appear to retain the drug and are 12.8 nm in diameter with an 8.0-nm pitch. Two classes of drugs have been distinguished: (i) those that bind to RecA protein and induce assembly at low relative concentrations (e.g., ethidium bromide) and (ii) those that do not appear to interact directly with RecA protein and must be present at relatively high drug concentrations to stimulate assembly (e.g., m-AMSA). Ethidium bromide, acridine orange, and quinacrine inhibit RecA protein binding to single-stranded DNA. Addition of ATP to the drug-induced filaments causes the protein to rapidly dissociate from dsDNA, and protein binding to dsDNA diminishes upon extended exposure to room light. We suggest that the structure of the drug-induced filaments may be more typical of the complex that initiates RecA protein assembly along DNA rather than the product of extensive polymerization as induced by adenosine 5'-[gamma-thio]triphosphate.

摘要

溴化乙锭、吖啶橙、4'-（9-吖啶基氨基）甲磺酰基邻茴香胺（o-AMSA）和间-AMSA可诱导RecA蛋白与双链（ds）DNA快速结合。形成的细丝似乎保留了药物，直径为12.8纳米，螺距为8.0纳米。已区分出两类药物：（i）那些在相对低浓度下与RecA蛋白结合并诱导组装的药物（例如溴化乙锭），以及（ii）那些似乎不直接与RecA蛋白相互作用且必须以相对高的药物浓度存在才能刺激组装的药物（例如间-AMSA）。溴化乙锭、吖啶橙和奎纳克林抑制RecA蛋白与单链DNA的结合。向药物诱导的细丝中添加ATP会导致蛋白质迅速从dsDNA上解离，并且在长时间暴露于室内光线下时，蛋白质与dsDNA的结合会减少。我们认为，药物诱导的细丝结构可能更典型地代表了沿DNA启动RecA蛋白组装的复合物，而不是由腺苷5'-[γ-硫代]三磷酸诱导的广泛聚合产物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b46/54260/fd179000e04a/pnas01038-0172-e.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b46/54260/9c211cfefcdb/pnas01038-0172-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b46/54260/7af5e2bf0159/pnas01038-0172-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b46/54260/0659c6e8938b/pnas01038-0172-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b46/54260/20430b104b38/pnas01038-0172-d.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b46/54260/fd179000e04a/pnas01038-0172-e.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b46/54260/9c211cfefcdb/pnas01038-0172-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b46/54260/7af5e2bf0159/pnas01038-0172-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b46/54260/0659c6e8938b/pnas01038-0172-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b46/54260/20430b104b38/pnas01038-0172-d.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b46/54260/fd179000e04a/pnas01038-0172-e.jpg

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本文引用的文献

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Measurement of protein-binding phenomena by gel filtration.通过凝胶过滤法测定蛋白质结合现象。
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Binding of the recA protein of Escherichia coli to single- and double-stranded DNA.大肠杆菌RecA蛋白与单链和双链DNA的结合。
J Biol Chem. 1981 Aug 25;256(16):8835-44.
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Unwinding of duplex DNA in complexes with recA protein.双链DNA与RecA蛋白形成复合物时的解旋。
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