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功能性哺乳动物U4小核核糖核蛋白的重组：Sm蛋白结合对于体外剪接并非必不可少。

Reconstitution of functional mammalian U4 small nuclear ribonucleoprotein: Sm protein binding is not essential for splicing in vitro.

作者信息

Wersig C, Bindereif A

机构信息

Max-Planck-Institut für Molekulare Genetik, Otto-Warburg-Laboratorium, Berlin, Germany.

出版信息

Mol Cell Biol. 1992 Apr;12(4):1460-8. doi: 10.1128/mcb.12.4.1460-1468.1992.

DOI:10.1128/mcb.12.4.1460-1468.1992
PMID:1532228
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC369587/
Abstract

We have developed an in vitro splicing complementation assay to investigate the domain structure of the mammalian U4 small nuclear RNA (snRNA) through mutational analysis. The addition of affinity-purified U4 snRNP or U4 RNA to U4-depleted nuclear extract efficiently restores splicing activity. In the U4-U6 interaction domain of U4 RNA, only stem II was found to be essential for splicing activity; the 5' loop is important for spliceosome stability. In the central domain, we have identified a U4 RNA sequence element that is important for splicing and spliceosome assembly. Surprisingly, an intact Sm domain is not essential for splicing in vitro. Our data provide evidence that several distinct regions of U4 RNA contribute to snRNP assembly, spliceosome assembly and stability, and splicing activity.

摘要

我们开发了一种体外剪接互补分析方法，通过突变分析来研究哺乳动物U4小核RNA（snRNA）的结构域。向去除U4的核提取物中添加亲和纯化的U4 snRNP或U4 RNA可有效恢复剪接活性。在U4 RNA的U4-U6相互作用结构域中，仅发现茎II对剪接活性至关重要；5'环对剪接体稳定性很重要。在中央结构域中，我们鉴定出一个对剪接和剪接体组装很重要的U4 RNA序列元件。令人惊讶的是，完整的Sm结构域对于体外剪接并非必不可少。我们的数据提供了证据，表明U4 RNA的几个不同区域有助于snRNP组装、剪接体组装和稳定性以及剪接活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c045/369587/afbcabd9bfa4/molcellb00168-0073-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c045/369587/080fc027bc90/molcellb00168-0070-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c045/369587/f66419952065/molcellb00168-0071-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c045/369587/4493dc1dc9ab/molcellb00168-0072-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c045/369587/14c736c07245/molcellb00168-0072-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c045/369587/1638e6e0f80f/molcellb00168-0073-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c045/369587/afbcabd9bfa4/molcellb00168-0073-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c045/369587/080fc027bc90/molcellb00168-0070-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c045/369587/f66419952065/molcellb00168-0071-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c045/369587/4493dc1dc9ab/molcellb00168-0072-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c045/369587/14c736c07245/molcellb00168-0072-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c045/369587/1638e6e0f80f/molcellb00168-0073-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c045/369587/afbcabd9bfa4/molcellb00168-0073-b.jpg

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剪接体激活：U4 是道路，茎环 I 是目标，Prp8 是守门员。让我们为 ATP 酶 Brr2 欢呼！
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