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U1小核核糖核蛋白颗粒特异性蛋白与U1 RNA的第一和第二个茎环相互作用，其中A蛋白独立于70K和Sm蛋白直接与RNA结合。

U1 small nuclear ribonucleoprotein particle-specific proteins interact with the first and second stem-loops of U1 RNA, with the A protein binding directly to the RNA independently of the 70K and Sm proteins.

作者信息

Patton J R, Habets W, van Venrooij W J, Pederson T

机构信息

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

出版信息

Mol Cell Biol. 1989 Aug;9(8):3360-8. doi: 10.1128/mcb.9.8.3360-3368.1989.

DOI:10.1128/mcb.9.8.3360-3368.1989
PMID:2529425
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC362381/
Abstract

The U1 small nuclear ribonucleoprotein particle (U1 snRNP), a cofactor in pre-mRNA splicing, contains three proteins, termed 70K, A, and C, that are not present in the other spliceosome-associated snRNPs. We studied the binding of the A and C proteins to U1 RNA, using a U1 snRNP reconstitution system and an antibody-induced nuclease protection technique. Antibodies that reacted with the A and C proteins induced nuclease protection of the first two stem-loops of U1 RNA in reconstituted U1 snRNP. Detailed analysis of the antibody-induced nuclease protection patterns indicated the existence of relatively long-range protein-protein interactions in the U1 snRNP, with the 5' end of U1 RNA and its associated specific proteins interacting with proteins bound to the Sm domain near the 3' end. UV cross-linking experiments in conjunction with an A-protein-specific antibody demonstrated that the A protein bound directly to the U1 RNA rather than assembling in the U1 snRNP exclusively via protein-protein interactions. This conclusion was supported by additional experiments revealing that the A protein could bind to U1 RNA in the absence of bound 70K and Sm core proteins.

摘要

U1小核核糖核蛋白颗粒（U1 snRNP）是前体mRNA剪接中的一种辅助因子，包含三种蛋白质，分别称为70K、A和C，而其他与剪接体相关的snRNP中不存在这些蛋白质。我们使用U1 snRNP重组系统和抗体诱导的核酸酶保护技术，研究了A蛋白和C蛋白与U1 RNA的结合。与A蛋白和C蛋白反应的抗体在重组的U1 snRNP中诱导了对U1 RNA前两个茎环的核酸酶保护。对抗体诱导的核酸酶保护模式的详细分析表明，U1 snRNP中存在相对远距离的蛋白质-蛋白质相互作用，U1 RNA的5'端及其相关的特定蛋白质与3'端附近与Sm结构域结合的蛋白质相互作用。结合A蛋白特异性抗体的紫外线交联实验表明，A蛋白直接与U1 RNA结合，而不是仅通过蛋白质-蛋白质相互作用组装到U1 snRNP中。其他实验表明，在没有结合的70K和Sm核心蛋白的情况下，A蛋白也能与U1 RNA结合，这一结论得到了这些实验的支持。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ade/362381/29df44df636a/molcellb00056-0222-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ade/362381/89e214410049/molcellb00056-0218-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ade/362381/b7e45bedd63c/molcellb00056-0219-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ade/362381/70772f240110/molcellb00056-0219-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ade/362381/fe5090136fd2/molcellb00056-0220-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ade/362381/00422fc0ad8c/molcellb00056-0221-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ade/362381/29df44df636a/molcellb00056-0222-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ade/362381/89e214410049/molcellb00056-0218-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ade/362381/b7e45bedd63c/molcellb00056-0219-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ade/362381/70772f240110/molcellb00056-0219-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ade/362381/fe5090136fd2/molcellb00056-0220-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ade/362381/00422fc0ad8c/molcellb00056-0221-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ade/362381/29df44df636a/molcellb00056-0222-a.jpg

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1
U1 small nuclear ribonucleoprotein particle-specific proteins interact with the first and second stem-loops of U1 RNA, with the A protein binding directly to the RNA independently of the 70K and Sm proteins.U1小核核糖核蛋白颗粒特异性蛋白与U1 RNA的第一和第二个茎环相互作用，其中A蛋白独立于70K和Sm蛋白直接与RNA结合。
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Nuclear ribonucleoprotein particles probed in living cells.在活细胞中检测的核糖核蛋白颗粒
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Monoclonal antibodies to nucleic acid-containing cellular constituents: probes for molecular biology and autoimmune disease.针对含核酸细胞成分的单克隆抗体：分子生物学和自身免疫性疾病的探针
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Multiple pseudouridine synthase activities for small nuclear RNAs.小核RNA的多种假尿苷合酶活性
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A pre-export U1 snRNP in Xenopus laevis oocyte nuclei.非洲爪蟾卵母细胞核中的一种预输出U1小核核糖核蛋白颗粒
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Pseudouridine formation in U2 small nuclear RNA.U2小核RNA中的假尿苷形成。
Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3324-8. doi: 10.1073/pnas.91.8.3324.
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Assembly, nuclear import and function of U7 snRNPs studied by microinjection of synthetic U7 RNA into Xenopus oocytes.通过将合成的U7 RNA显微注射到非洲爪蟾卵母细胞中来研究U7小核核糖核蛋白颗粒的组装、核输入及功能。
Nucleic Acids Res. 1995 Aug 25;23(16):3141-51. doi: 10.1093/nar/23.16.3141.
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Identification of the RNA binding segment of human U1 A protein and definition of its binding site on U1 snRNA.
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腺病毒感染和未感染的HeLa细胞中SS-B（La）抗原的特性分析。
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Clin Exp Immunol. 1983 Oct;54(1):265-76.
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The 5' terminus of the RNA moiety of U1 small nuclear ribonucleoprotein particles is required for the splicing of messenger RNA precursors.U1小核核糖核蛋白颗粒的RNA部分的5'末端是信使RNA前体剪接所必需的。
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The structure of mammalian small nuclear ribonucleoproteins. Identification of multiple protein components reactive with anti-(U1)ribonucleoprotein and anti-Sm autoantibodies.哺乳动物小核核糖核蛋白的结构。与抗（U1）核糖核蛋白和抗Sm自身抗体反应的多种蛋白质成分的鉴定。
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Characterization of U small nuclear RNA-associated proteins.U小核RNA相关蛋白的特性分析
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Characterization of the autoimmune antigenic determinant for ribonucleoprotein (RNP) antibody.核糖核蛋白（RNP）抗体自身免疫抗原决定簇的特征分析。
Immunol Commun. 1984;13(2):137-49. doi: 10.3109/08820138409025457.
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The U1 small nuclear RNA-protein complex selectively binds a 5' splice site in vitro.U1小核核糖核蛋白复合体在体外能选择性结合5'剪接位点。
Cell. 1983 Jun;33(2):509-18. doi: 10.1016/0092-8674(83)90432-4.