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前体信使核糖核酸在离散核位点的定位。

Localization of pre-messenger RNA at discrete nuclear sites.

作者信息

Wang J, Cao L G, Wang Y L, Pederson T

机构信息

Cell Biology Group, Worcester Foundation for Experimental Biology, Shrewsbury, MA 01545.

出版信息

Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7391-5. doi: 10.1073/pnas.88.16.7391.

DOI:10.1073/pnas.88.16.7391
PMID:1831271
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC52301/
Abstract

We have studied the nuclear localization of rhodamine-labeled pre-mRNA after microinjection into nuclei of cultured rat kidney epithelial cells. Intranuclear localization of the injected RNA was followed in the living cells by fluorescence microscopy and digital image processing. Injected human beta-globin pre-mRNA became localized in 30-60 discrete nuclear sites that were coincident with loci defined by monoclonal antibodies against small nuclear ribonucleoproteins (Sm) or another spliceosome component (SC-35) in parallel immunocytochemical studies on the same nuclei. Similar patterns of nuclear localization were observed with a rat proenkephalin pre-mRNA. Nuclear microinjection of an intronlacking beta-globin RNA, a splicing-defective beta-globin mutant pre-mRNA, or an antisense beta-globin pre-mRNA did not result in localization at discrete sites. These results indicate that pre-mRNA binds preferentially to nuclear Sm and SC-35 antibody-reactive sites in vivo and that the binding requires intron sequences.

摘要

我们研究了将罗丹明标记的前体mRNA显微注射到培养的大鼠肾上皮细胞核中后的核定位情况。通过荧光显微镜和数字图像处理在活细胞中追踪注射RNA的核内定位。在对相同细胞核进行的平行免疫细胞化学研究中，注射的人β-珠蛋白前体mRNA定位于30 - 60个离散的核位点，这些位点与针对小核核糖核蛋白（Sm）或另一种剪接体成分（SC-35）的单克隆抗体所定义的位点一致。用大鼠前脑啡肽原前体mRNA观察到了类似的核定位模式。对缺乏内含子的β-珠蛋白RNA、剪接缺陷型β-珠蛋白突变体前体mRNA或反义β-珠蛋白前体mRNA进行核显微注射，并未导致其在离散位点定位。这些结果表明，前体mRNA在体内优先与核Sm和SC-35抗体反应位点结合，且这种结合需要内含子序列。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dae4/52301/eaab50f3c644/pnas01066-0504-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dae4/52301/68367fdc7037/pnas01066-0502-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dae4/52301/169cfe70a00e/pnas01066-0502-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dae4/52301/8e0235927f33/pnas01066-0502-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dae4/52301/792f2ce07d1f/pnas01066-0503-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dae4/52301/5641cc75d414/pnas01066-0503-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dae4/52301/eaab50f3c644/pnas01066-0504-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dae4/52301/68367fdc7037/pnas01066-0502-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dae4/52301/169cfe70a00e/pnas01066-0502-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dae4/52301/8e0235927f33/pnas01066-0502-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dae4/52301/792f2ce07d1f/pnas01066-0503-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dae4/52301/5641cc75d414/pnas01066-0503-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dae4/52301/eaab50f3c644/pnas01066-0504-a.jpg

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本文引用的文献

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Human beta-globin pre-mRNA synthesized in vitro is accurately spliced in Xenopus oocyte nuclei.在体外合成的人β-珠蛋白前体信使核糖核酸（pre-mRNA）在非洲爪蟾卵母细胞核中能被精确剪接。
Cell. 1983 Mar;32(3):681-94. doi: 10.1016/0092-8674(83)90054-5.
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Normal and mutant human beta-globin pre-mRNAs are faithfully and efficiently spliced in vitro.正常和突变的人β-珠蛋白前体mRNA在体外能够被准确且高效地剪接。
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Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes.
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Cue-Polarized Transport of β-actin mRNA Depends on 3'UTR and Microtubules in Live Growth Cones.β-肌动蛋白mRNA的线索极化运输取决于活生长锥中的3'非翻译区和微管。
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