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由SV40大T抗原催化的不依赖序列的双链DNA解旋反应。

Sequence independent duplex DNA opening reaction catalysed by SV40 large tumor antigen.

作者信息

Scheffner M, Wessel R, Stahl H

机构信息

Fakultät für Biologie, Universität Konstanz, FRG.

出版信息

Nucleic Acids Res. 1989 Jan 11;17(1):93-106. doi: 10.1093/nar/17.1.93.

DOI:10.1093/nar/17.1.93
PMID:2536153
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC331537/
Abstract

Simian virus 40 (SV40) large tumor antigen (T antigen) is mainly localized in the nucleus where it exhibits two biochemical properties: DNA binding and helicase activity. Both activities are necessary for viral DNA replication and may also enable T antigen to modulate cellular growth. Here we present biochemical and electron microscopic evidence that the helicase activity can start at internal sites of fully double-stranded DNA molecules not containing the SV40 origin or replication. Using T antigen specific monoclonal antibodies, this unwinding reaction can be biochemically divided in an initiation (duplex opening) and a propagation step. The duplex opening reaction (as well as the propagation step) does not depend on a specific DNA sequence or secondary structure. In addition, we have found that T antigen forms an ATP dependent nucleoprotein complex at double-stranded DNA, which may be an essential step for the sequence independent duplex DNA opening reaction.

摘要

猴病毒40（SV40）大T抗原主要定位于细胞核，在细胞核中它表现出两种生化特性：DNA结合和螺旋酶活性。这两种活性对于病毒DNA复制都是必需的，也可能使T抗原调节细胞生长。在此，我们提供了生化和电子显微镜证据，表明螺旋酶活性可以在不包含SV40复制起点的完全双链DNA分子的内部位点起始。使用T抗原特异性单克隆抗体，这种解旋反应在生化上可分为起始（双链打开）和延伸步骤。双链打开反应（以及延伸步骤）不依赖于特定的DNA序列或二级结构。此外，我们发现T抗原在双链DNA上形成一种依赖ATP的核蛋白复合物，这可能是序列非依赖性双链DNA打开反应的一个关键步骤。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ec3f/331537/3c35000cad30/nar00210-0117-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ec3f/331537/af113d4bc4f6/nar00210-0112-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ec3f/331537/d22b770639f4/nar00210-0113-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ec3f/331537/c6a0f9bd5518/nar00210-0115-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ec3f/331537/6e7823314f30/nar00210-0116-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ec3f/331537/3c35000cad30/nar00210-0117-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ec3f/331537/af113d4bc4f6/nar00210-0112-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ec3f/331537/d22b770639f4/nar00210-0113-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ec3f/331537/c6a0f9bd5518/nar00210-0115-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ec3f/331537/6e7823314f30/nar00210-0116-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ec3f/331537/3c35000cad30/nar00210-0117-a.jpg

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本文引用的文献

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Simplified preparation of rabbit Fab fragments.兔Fab片段的简易制备方法。
J Immunol Methods. 1983 Apr 29;59(2):199-203. doi: 10.1016/0022-1759(83)90031-5.
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Essential contact residues within SV40 large T antigen binding sites I and II identified by alkylation-interference.通过烷基化干扰鉴定出的SV40大T抗原结合位点I和II内的必需接触残基。
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