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与U5 snRNP结合的反义寡核苷酸会诱导构象变化，从而暴露U5 snRNA的保守环。

Antisense oligonucleotide binding to U5 snRNP induces a conformational change that exposes the conserved loop of U5 snRNA.

作者信息

Ast G, Weiner A M

机构信息

Department of Molecular Biophysics and Biochemistry, Yale University, 266 Whitney Avenue, PO Box 208114, New Haven, CT 06520-8114, USA.

出版信息

Nucleic Acids Res. 1997 Sep 1;25(17):3508-13. doi: 10.1093/nar/25.17.3508.

DOI:10.1093/nar/25.17.3508
PMID:9254712
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC146904/
Abstract

Conformational rearrangements of the spliceosomal small nuclear RNAs (U snRNAs) are essential for proper assembly of the active site prior to the first catalytic step of splicing. We have previously shown that conformational changes caused by binding of an antisense 2'-O-methyl RNA oligonucleotide (BU5Ae) to U5 snRNA nt 68-88 disrupted the U4/U5/U6 complex and induced formation of the U1/U4/U5 and U2/U6 complexes. Here we show that the conformational change induced by BU5Ae exposes the invariant loop of U5 that binds the 5'exon and also reorganizes internal loop 1 (IL1) and the top of stem 2. Interestingly, we have also previously found that the U1/U4/U5 complex induced by BU5Ae brings the invariant loop of U5 into close proximity with the 5'-end of U1. Taken together, these data suggest that U1 and U5 may both contribute to the ability of the U1/U4/U5 complex to bind the 5' splice site.

摘要

剪接体小核RNA（U snRNAs）的构象重排对于剪接第一步催化反应之前活性位点的正确组装至关重要。我们之前已经表明，反义2'-O-甲基RNA寡核苷酸（BU5Ae）与U5 snRNA nt 68-88结合所引起的构象变化破坏了U4/U5/U6复合物，并诱导形成了U1/U4/U5和U2/U6复合物。在此我们表明，BU5Ae诱导的构象变化暴露了U5与5'外显子结合的恒定环，并且还重新组织了内部环1（IL1）和茎2的顶部。有趣的是，我们之前还发现，由BU5Ae诱导的U1/U4/U5复合物使U5的恒定环与U1的5'末端紧密靠近。综上所述，这些数据表明U1和U5可能都有助于U1/U4/U5复合物结合5'剪接位点的能力。

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本文引用的文献

1
A novel U1/U5 interaction indicates proximity between U1 and U5 snRNAs during an early step of mRNA splicing.一种新型的U1/U5相互作用表明，在mRNA剪接的早期步骤中，U1和U5小核RNA（snRNA）之间存在空间上的接近。
RNA. 1997 Apr;3(4):371-81.
2
A spontaneous duplication in U6 spliceosomal RNA uncouples the early and late functions of the ACAGA element in vivo.U6剪接体RNA中的自发重复在体内使ACAGA元件的早期和晚期功能解偶联。
RNA. 1996 Sep;2(9):879-94.
3
The invariant U5 snRNA loop 1 sequence is dispensable for the first catalytic step of pre-mRNA splicing in yeast.不变的U5小核仁RNA环1序列对于酵母中前体mRNA剪接的第一步催化反应并非必需。
Cell. 1996 Aug 23;86(4):679-89. doi: 10.1016/s0092-8674(00)80140-3.
4
A U1/U4/U5 snRNP complex induced by a 2'-O-methyl-oligoribonucleotide complementary to U5 snRNA.一种由与U5 snRNA互补的2'-O-甲基寡核糖核苷酸诱导形成的U1/U4/U5 snRNP复合物。
Science. 1996 May 10;272(5263):881-4. doi: 10.1126/science.272.5263.881.
5
The canonical GU dinucleotide at the 5' splice site is recognized by p220 of the U5 snRNP within the spliceosome.剪接体内U5小核核糖核蛋白（snRNP）的p220可识别位于5'剪接位点的典型GU二核苷酸。
RNA. 1996 Mar;2(3):213-25.
6
snRNA interactions at 5' and 3' splice sites monitored by photoactivated crosslinking in yeast spliceosomes.通过酵母剪接体中的光活化交联监测5'和3'剪接位点处的小核RNA相互作用。
RNA. 1995 Nov;1(9):968-80.
7
A functional association between the 5' and 3' splice site is established in the earliest prespliceosome complex (E) in mammals.在哺乳动物最早的前剪接体复合物（E）中，5'和3'剪接位点之间建立了功能关联。
Genes Dev. 1993 Jun;7(6):1008-20. doi: 10.1101/gad.7.6.1008.
8
Involvement of U6 snRNA in 5' splice site selection.U6小核RNA参与5'剪接位点的选择。
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9
The U5 and U6 small nuclear RNAs as active site components of the spliceosome.U5和U6小核RNA作为剪接体的活性位点组分。
Science. 1993 Dec 24;262(5142):1989-96. doi: 10.1126/science.8266094.
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Mutations in U6 snRNA that alter splice site specificity: implications for the active site.改变剪接位点特异性的U6小核仁RNA突变：对活性位点的影响
Science. 1993 Dec 24;262(5142):1982-8. doi: 10.1126/science.8266093.