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差异性核酸酶敏感性鉴定出酵母前体mRNA与剪接体之间的紧密接触。

Differential nuclease sensitivity identifies tight contacts between yeast pre-mRNA and spliceosomes.

作者信息

Rymond B C, Rosbash M

出版信息

EMBO J. 1986 Dec 20;5(13):3517-23. doi: 10.1002/j.1460-2075.1986.tb04677.x.

DOI:10.1002/j.1460-2075.1986.tb04677.x
PMID:2435546
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1167388/
Abstract

The oligonucleotide-directed RNase H sensitivity of a yeast (Saccharomyces cerevisiae) pre-mRNA was determined in an in vitro splicing reaction. While most of the pre-mRNA was sensitive to cleavage, the regions of the 5' splice site and TACTAAC box were found to be highly resistant. The biochemical requirements for protection against nuclease attack parallel those of both spliceosome formation and splicing. Most of the uncleaved pre-mRNA remaining after RNase H challenge was found associated with two forms of the yeast spliceosome. Differences in the RNase H sensitivity of pre-mRNA found in the two spliceosome forms indicate an increased association of splicing factors with the 5' splice site during spliceosome assembly.

摘要

在体外剪接反应中测定了酵母（酿酒酵母）前体mRNA的寡核苷酸定向核糖核酸酶H敏感性。虽然大多数前体mRNA对切割敏感，但发现5'剪接位点和TACTAAC框区域具有高度抗性。防止核酸酶攻击的生化要求与剪接体形成和剪接的要求相似。核糖核酸酶H攻击后剩余的大多数未切割前体mRNA被发现与酵母剪接体的两种形式相关。在两种剪接体形式中发现的前体mRNA核糖核酸酶H敏感性差异表明，在剪接体组装过程中，剪接因子与5'剪接位点的结合增加。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1038/1167388/bc438aa2ae8c/emboj00176-0121-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1038/1167388/0f051bbef4da/emboj00176-0118-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1038/1167388/ca620486370a/emboj00176-0119-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1038/1167388/021380faf76a/emboj00176-0119-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1038/1167388/5c6a42261de5/emboj00176-0120-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1038/1167388/222d5daf7787/emboj00176-0121-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1038/1167388/bc438aa2ae8c/emboj00176-0121-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1038/1167388/0f051bbef4da/emboj00176-0118-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1038/1167388/ca620486370a/emboj00176-0119-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1038/1167388/021380faf76a/emboj00176-0119-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1038/1167388/5c6a42261de5/emboj00176-0120-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1038/1167388/222d5daf7787/emboj00176-0121-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1038/1167388/bc438aa2ae8c/emboj00176-0121-b.jpg

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1
Differential nuclease sensitivity identifies tight contacts between yeast pre-mRNA and spliceosomes.差异性核酸酶敏感性鉴定出酵母前体mRNA与剪接体之间的紧密接触。
EMBO J. 1986 Dec 20;5(13):3517-23. doi: 10.1002/j.1460-2075.1986.tb04677.x.
2
Identification of sites of pre-MRNA/spliceosome association.前体mRNA/剪接体结合位点的鉴定
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3
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4
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Alterations of RNase H sensitivity of the 3' splice site region during the in vitro splicing reaction.体外剪接反应过程中3'剪接位点区域核糖核酸酶H敏感性的改变。
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A chemical modification/interference study of yeast pre-mRNA spliceosome assembly and splicing.酵母前体信使核糖核酸剪接体组装与剪接的化学修饰/干扰研究
Genes Dev. 1988 Apr;2(4):428-39. doi: 10.1101/gad.2.4.428.
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The "spliceosome": yeast pre-messenger RNA associates with a 40S complex in a splicing-dependent reaction.“剪接体”：酵母前体信使核糖核酸在剪接依赖性反应中与一个40S复合体结合。
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Yeast mRNA splicing in vitro.体外酵母mRNA剪接
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Specific accessory sequences in Saccharomyces cerevisiae introns control assembly of pre-mRNAs into spliceosomes.酿酒酵母内含子中的特定辅助序列控制前体mRNA组装成剪接体。
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10
Multiple interactions between the splicing substrate and small nuclear ribonucleoproteins in spliceosomes.剪接底物与剪接体中的小核核糖核蛋白之间的多种相互作用。
Mol Cell Biol. 1987 Jan;7(1):281-93. doi: 10.1128/mcb.7.1.281-293.1987.

引用本文的文献

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Invariant U2 snRNA nucleotides form a stem loop to recognize the intron early in splicing.不变 U2 snRNA 核苷酸形成茎环结构以在剪接早期识别内含子。
Mol Cell. 2010 May 14;38(3):416-27. doi: 10.1016/j.molcel.2010.02.036.
2
Pathways for selection of 5' splice sites by U1 snRNPs and SF2/ASF.U1 小核核糖核蛋白颗粒（snRNPs）和 SF2/ASF 选择 5' 剪接位点的途径。
EMBO J. 1993 Sep;12(9):3607-17. doi: 10.1002/j.1460-2075.1993.tb06034.x.
3
Interaction of the yeast splicing factor PRP8 with substrate RNA during both steps of splicing.酵母剪接因子PRP8在剪接的两个步骤中与底物RNA的相互作用。

本文引用的文献

1
Evidence for the biochemical role of an internal sequence in yeast nuclear mRNA introns: implications for U1 RNA and metazoan mRNA splicing.酵母核mRNA内含子内部序列的生化作用证据：对U1 RNA和后生动物mRNA剪接的影响
Cell. 1983 Sep;34(2):395-403. doi: 10.1016/0092-8674(83)90373-2.
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A comparison of yeast ribosomal protein gene DNA sequences.酵母核糖体蛋白基因DNA序列的比较
Nucleic Acids Res. 1984 Nov 26;12(22):8295-312. doi: 10.1093/nar/12.22.8295.
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Point mutations identify the conserved, intron-contained TACTAAC box as an essential splicing signal sequence in yeast.
Nucleic Acids Res. 1995 Feb 11;23(3):320-6. doi: 10.1093/nar/23.3.320.
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Two spliceosomes can form simultaneously and independently on synthetic double-intron messenger RNA precursors.两个剪接体可以在合成的双内含子信使核糖核酸前体上同时且独立地形成。
EMBO J. 1987 Jun;6(6):1747-55. doi: 10.1002/j.1460-2075.1987.tb02427.x.
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Mol Cell Biol. 1988 Sep;8(9):3755-60. doi: 10.1128/mcb.8.9.3755-3760.1988.
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The U1 RNA-binding site of the U1 small nuclear ribonucleoprotein (snRNP)-associated A protein suggests a similarity with U2 snRNPs.与U1小核核糖核蛋白（snRNP）相关的A蛋白的U1 RNA结合位点表明其与U2 snRNPs存在相似性。
Mol Cell Biol. 1989 Jul;9(7):2975-82. doi: 10.1128/mcb.9.7.2975-2982.1989.
7
The PRP4 (RNA4) protein of Saccharomyces cerevisiae is associated with the 5' portion of the U4 small nuclear RNA.酿酒酵母的PRP4（RNA4）蛋白与U4小核RNA的5'部分相关联。
Mol Cell Biol. 1990 Mar;10(3):1217-25. doi: 10.1128/mcb.10.3.1217-1225.1990.
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Protein interactions in nuclear pre-mRNA splicing in Saccharomyces cerevisiae.酿酒酵母中核内前体mRNA剪接过程中的蛋白质相互作用
Mol Biol Rep. 1990;14(2-3):141-2. doi: 10.1007/BF00360449.
9
A nuclear micrococcal-sensitive, ATP-dependent exoribonuclease degrades uncapped but not capped RNA substrates.一种对核微球菌敏感、依赖ATP的外切核糖核酸酶可降解无帽RNA底物，但不能降解有帽RNA底物。
Nucleic Acids Res. 1991 May 25;19(10):2685-92. doi: 10.1093/nar/19.10.2685.
10
Small sequence insertions within the branch point region dictate alternative sites of lariat formation in a yeast intron.分支点区域内的小序列插入决定了酵母内含子中套索形成的替代位点。
Nucleic Acids Res. 1992 Dec 25;20(24):6649-55. doi: 10.1093/nar/20.24.6649.
点突变确定了保守的、内含于内含子中的TACTAAC盒是酵母中一个必需的剪接信号序列。
Cell. 1984 Mar;36(3):645-53. doi: 10.1016/0092-8674(84)90344-1.
4
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。
Cell. 1984 Apr;36(4):993-1005. doi: 10.1016/0092-8674(84)90049-7.
5
Expression of a beta-galactosidase gene containing the ribosomal protein 51 intron is sensitive to the rna2 mutation of yeast.含有核糖体蛋白51内含子的β-半乳糖苷酶基因的表达对酵母的rna2突变敏感。
Proc Natl Acad Sci U S A. 1983 Jul;80(14):4403-7. doi: 10.1073/pnas.80.14.4403.
6
Evidence for an intron-contained sequence required for the splicing of yeast RNA polymerase II transcripts.酵母RNA聚合酶II转录本剪接所需的内含子包含序列的证据。
Cell. 1983 Jun;33(2):519-27. doi: 10.1016/0092-8674(83)90433-6.
7
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.
8
Role of the 3' splice site consensus sequence in mammalian pre-mRNA splicing.3'剪接位点共有序列在哺乳动物前体mRNA剪接中的作用。
Nature. 1985;317(6039):732-4. doi: 10.1038/317732a0.
9
A quantitative analysis of the effects of 5' junction and TACTAAC box mutants and mutant combinations on yeast mRNA splicing.5' 连接区和TACTAAC盒突变体及突变体组合对酵母mRNA剪接影响的定量分析。
Cell. 1985 Dec;43(2 Pt 1):423-30. doi: 10.1016/0092-8674(85)90172-2.
10
Cleavage of 5' splice site and lariat formation are independent of 3' splice site in yeast mRNA splicing.在酵母mRNA剪接过程中，5'剪接位点的切割和套索结构的形成与3'剪接位点无关。
Nature. 1985;317(6039):735-7. doi: 10.1038/317735a0.