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三链螺旋形成寡核苷酸与病毒RNA对HIV-1逆转录的抑制作用。

Inhibition of HIV-1 reverse transcription by triple-helix forming oligonucleotides with viral RNA.

作者信息

Volkmann S, Jendis J, Frauendorf A, Moelling K

机构信息

Max-Planck-Institut für Molekulare Genetik, Abt. Schuster, Berlin Dahlem, Germany.

出版信息

Nucleic Acids Res. 1995 Apr 11;23(7):1204-12. doi: 10.1093/nar/23.7.1204.

DOI:10.1093/nar/23.7.1204
PMID:7537875
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC306832/
Abstract

Reverse transcription of retroviral RNA into double-stranded DNA is catalyzed by reverse transcriptase (RT). A highly conserved polypurine tract (PPT) on the viral RNA serves as primer for plus-strand DNA synthesis and is a possible target for triple-helix formation. Triple-helix formation during reverse transcription involves either single-stranded RNA or an RNA.DNA hybrid. The effect of triple-helix formation on reverse transcription has been analyzed here in vitro using a three-strand-system consisting of an RNA.DNA hybrid and triplex-forming oligonucleotides (TFOs) consisting either of DNA or RNA. Three strand triple-helices inhibit RNase H cleavage of the PPT-RNA.DNA hybrid and initiation of plus-strand DNA synthesis in vitro. Triple-helix formation on a single-stranded RNA target has also been tested in a two-strand-system with TFOs comprising Watson-Crick and Hoogsteen base-pairing sequences, both targeted to the PPT-RNA, on a single strand connected by a linker (T)4. TFOs prevent RNase H cleavage of the PPT-RNA and initiation of plus-strand DNA synthesis in vitro. In cell culture experiments one TFO is an efficient inhibitor of retrovirus replication, leading to a block of p24 synthesis and inhibition of syncytia formation in newly infected cells.

摘要

逆转录病毒RNA逆转录成双链DNA是由逆转录酶（RT）催化的。病毒RNA上一个高度保守的多聚嘌呤序列（PPT）作为正链DNA合成的引物，并且是三链体形成的一个可能靶点。逆转录过程中的三链体形成涉及单链RNA或RNA-DNA杂交体。本文在体外使用由RNA-DNA杂交体和由DNA或RNA组成的三链体形成寡核苷酸（TFO）构成的三链系统分析了三链体形成对逆转录的影响。三链三链体在体外抑制PPT-RNA-DNA杂交体的核糖核酸酶H切割以及正链DNA合成的起始。单链RNA靶点上的三链体形成也已在双链系统中进行了测试，该双链系统中的TFO包含通过接头（T）4连接在单链上的针对PPT-RNA的沃森-克里克碱基对和 hoogsteen碱基对序列。TFO在体外阻止PPT-RNA的核糖核酸酶H切割以及正链DNA合成的起始。在细胞培养实验中，一种TFO是逆转录病毒复制的有效抑制剂，导致新感染细胞中p24合成受阻并抑制合胞体形成。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6804/306832/17e7ea8bdd0c/nar00007-0129-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6804/306832/78fcd3889fda/nar00007-0127-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6804/306832/da672ff10dd6/nar00007-0128-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6804/306832/4b1715220575/nar00007-0129-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6804/306832/17e7ea8bdd0c/nar00007-0129-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6804/306832/78fcd3889fda/nar00007-0127-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6804/306832/da672ff10dd6/nar00007-0128-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6804/306832/4b1715220575/nar00007-0129-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6804/306832/17e7ea8bdd0c/nar00007-0129-b.jpg

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