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5,6-二氯-1-β-D-呋喃核糖基苯并咪唑（DRB）诱导小鼠β-珠蛋白基因转录提前终止

Induction of premature termination of transcription of the mouse beta-globin gene by 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB).

作者信息

Tweeten K A, Molloy G R

出版信息

Nucleic Acids Res. 1981 Jul 24;9(14):3307-19. doi: 10.1093/nar/9.14.3307.

DOI:10.1093/nar/9.14.3307
PMID:6269079
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC327353/
Abstract

Hybridization of pulse-labeled RNA from DRB-treated Friend cells to mouse beta-globin cDNA revealed that the appearance of beta-globin mRNA in the cytoplasm was inhibited by greater than 87%. To examine the effect of DRB (125 microM) on HnRNA synthesis, nuclear RNA was electrophoresed in methyl mercuric hydroxide gels, transferred to nitrocellulose, and hybridized with beta-globin specific probes. Full-length nuclear transcripts, while present in untreated cells, were not detected in DRB-treated cells. Using restriction enzymes, the cloned beta-globin gene was divided into fragments proceeding from the 5' gene region to the 3' gene region. RNA labeled in vitro by transcription in nuclei isolated from DRB-treated cells hybridized only to the promoter proximal DNA fragment. Transcripts hybridizing to fragments from both the 5' and 3' regions of the gene were produced in nuclei from untreated cells. Together these results indicate that DRB causes premature termination of transcription within the beta-globin gene.

摘要

用脉冲标记的来自经DRB处理的Friend细胞的RNA与小鼠β-珠蛋白cDNA杂交，结果显示β-珠蛋白mRNA在细胞质中的出现受到了超过87%的抑制。为了检测DRB（125微摩尔）对核不均一RNA（HnRNA）合成的影响，将核RNA在氢氧化甲基汞凝胶中进行电泳，转移至硝酸纤维素膜上，并用β-珠蛋白特异性探针进行杂交。全长核转录本在未处理的细胞中存在，但在经DRB处理的细胞中未检测到。使用限制性内切酶，将克隆的β-珠蛋白基因从5'基因区域到3'基因区域分成片段。在从经DRB处理的细胞中分离出的细胞核中通过体外转录标记的RNA仅与启动子近端DNA片段杂交。与基因5'和3'区域片段杂交的转录本在未处理细胞的细胞核中产生。这些结果共同表明，DRB导致β-珠蛋白基因内转录的过早终止。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61d3/327353/d8d2ee2224d3/nar00407-0103-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61d3/327353/e8211ffafe22/nar00407-0100-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61d3/327353/c30ad99c54a6/nar00407-0102-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61d3/327353/d8d2ee2224d3/nar00407-0103-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61d3/327353/e8211ffafe22/nar00407-0100-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61d3/327353/c30ad99c54a6/nar00407-0102-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61d3/327353/d8d2ee2224d3/nar00407-0103-a.jpg

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本文引用的文献

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Short capped hnRNA precursor chains in HeLa cells: continued synthesis in the presence of 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole.HeLa细胞中短帽hnRNA前体链：在5,6-二氯-1-β-D-呋喃核糖基苯并咪唑存在下的持续合成
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The addition of 5' cap structures occurs early in hnRNA synthesis and prematurely terminated molecules are capped.5'帽结构的添加在核内不均一RNA（hnRNA）合成早期发生，并且提前终止的分子也会被加帽。
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P-TEFb kinase is required for HIV Tat transcriptional activation in vivo and in vitro.P-TEFb激酶在体内和体外对于HIV Tat转录激活都是必需的。
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Minute virus of mice infection modifies cellular transcription elongation.小鼠微小病毒感染会改变细胞转录延伸。
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Mechanism of action of dichloro-beta-D-ribofuranosylbenzimidazole: effect on in vitro transcription.二氯-β-D-呋喃核糖基苯并咪唑的作用机制：对体外转录的影响
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