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体外mRNA剪接所需的来自HeLa细胞的两个组分的分离与特性鉴定。

Isolation and characterization of two fractions from HeLa cells required for mRNA splicing in vitro.

作者信息

Furneaux H M, Perkins K K, Freyer G A, Arenas J, Hurwitz J

出版信息

Proc Natl Acad Sci U S A. 1985 Jul;82(13):4351-5. doi: 10.1073/pnas.82.13.4351.

DOI:10.1073/pnas.82.13.4351
PMID:3859867
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC390411/
Abstract

A nuclear extract from HeLa cells has been separated by DEAE-cellulose chromatography into two fractions, both of which are required for mRNA splicing in vitro. Both fractions are heat labile and sensitive to N-ethylmaleimide. The activity of one of the fractions was abolished by preincubation with micrococcal nuclease, while the other fraction was unaffected by this treatment. This abolition indicates an essential nucleic acid component. Fractions I and II are required for the in vitro splicing of human beta-globin and adenovirus transcripts.

摘要

从HeLa细胞中提取的核提取物经DEAE-纤维素色谱法分离为两个组分，二者都是体外mRNA剪接所必需的。两个组分都对热不稳定且对N-乙基马来酰亚胺敏感。其中一个组分的活性在与微球菌核酸酶预孵育后被消除，而另一个组分不受此处理的影响。这种消除表明存在一种必需的核酸成分。组分I和组分II是人类β-珠蛋白和腺病毒转录本体外剪接所必需的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f79/390411/017e5de90073/pnas00353-0061-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f79/390411/4a6dbc9b420f/pnas00353-0059-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f79/390411/dd19b8e044cb/pnas00353-0059-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f79/390411/2417f489d763/pnas00353-0059-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f79/390411/c2db55ef2bea/pnas00353-0060-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f79/390411/faa7333634f5/pnas00353-0061-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f79/390411/017e5de90073/pnas00353-0061-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f79/390411/4a6dbc9b420f/pnas00353-0059-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f79/390411/dd19b8e044cb/pnas00353-0059-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f79/390411/2417f489d763/pnas00353-0059-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f79/390411/c2db55ef2bea/pnas00353-0060-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f79/390411/faa7333634f5/pnas00353-0061-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f79/390411/017e5de90073/pnas00353-0061-b.jpg

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Isolation and characterization of two fractions from HeLa cells required for mRNA splicing in vitro.体外mRNA剪接所需的来自HeLa细胞的两个组分的分离与特性鉴定。
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本文引用的文献

1
Are snRNPs involved in splicing?小核核糖核蛋白颗粒参与剪接过程吗？
Nature. 1980 Jan 10;283(5743):220-4. doi: 10.1038/283220a0.
2
Bacteriophage SP6-specific RNA polymerase. I. Isolation and characterization of the enzyme.噬菌体SP6特异性RNA聚合酶。I. 酶的分离与特性鉴定。
J Biol Chem. 1982 May 25;257(10):5772-8.
3
Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.从分离的哺乳动物细胞核的可溶性提取物中，RNA聚合酶II进行准确的转录起始。
前体信使核糖核酸剪接所需蛋白质的纯化及其与套索脱支酶的分离。
EMBO J. 1985 Dec 16;4(13A):3571-81. doi: 10.1002/j.1460-2075.1985.tb04119.x.
4
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.
5
Fractionation and characterization of a yeast mRNA splicing extract.酵母mRNA剪接提取物的分级分离与特性分析。
Proc Natl Acad Sci U S A. 1986 Apr;83(8):2387-91. doi: 10.1073/pnas.83.8.2387.
6
RNA splicing products formed with isolated fractions from HeLa cells are associated with fast-sedimenting complexes.由从HeLa细胞中分离出的组分形成的RNA剪接产物与快速沉降复合物相关。
Proc Natl Acad Sci U S A. 1986 Feb;83(4):887-91. doi: 10.1073/pnas.83.4.887.
7
Requirements for accurate and efficient mRNA 3' end cleavage and polyadenylation of a simian virus 40 early pre-RNA in vitro.
Mol Cell Biol. 1987 Jan;7(1):495-503. doi: 10.1128/mcb.7.1.495-503.1987.
8
Pre-mRNA splicing and the nuclear matrix.前体信使核糖核酸剪接与核基质。
Mol Cell Biol. 1987 Jan;7(1):111-20. doi: 10.1128/mcb.7.1.111-120.1987.
9
Three distinct activities possibly involved in mRNA splicing are found in a nuclear fraction lacking U1 and U2 RNA.在缺乏U1和U2 RNA的细胞核组分中发现了可能参与mRNA剪接的三种不同活性。
Nucleic Acids Res. 1986 Apr 11;14(7):3045-57. doi: 10.1093/nar/14.7.3045.
10
An ordered pathway of snRNP binding during mammalian pre-mRNA splicing complex assembly.哺乳动物前体mRNA剪接复合体组装过程中snRNP结合的有序途径。
EMBO J. 1987 Aug;6(8):2415-24. doi: 10.1002/j.1460-2075.1987.tb02520.x.
Nucleic Acids Res. 1983 Mar 11;11(5):1475-89. doi: 10.1093/nar/11.5.1475.
4
Cofactor requirements of splicing of purified messenger RNA precursors.纯化信使RNA前体剪接的辅因子需求
Nature. 1984;308(5957):375-7. doi: 10.1038/308375a0.
5
In vitro-synthesized adenovirus 2 messenger RNA precursors are accurately spliced by nuclear extracts.体外合成的腺病毒2信使核糖核酸前体被核提取物精确剪接。
Proc Natl Acad Sci U S A. 1984 Aug;81(15):4707-11. doi: 10.1073/pnas.81.15.4707.
6
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.
7
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。
Cell. 1984 Aug;38(1):299-307. doi: 10.1016/0092-8674(84)90551-8.
8
Lariat RNA's as intermediates and products in the splicing of messenger RNA precursors.套索RNA作为信使RNA前体剪接过程中的中间体和产物。
Science. 1984 Aug 31;225(4665):898-903. doi: 10.1126/science.6206566.
9
Splicing of in vitro synthesized messenger RNA precursors in HeLa cell extracts.体外合成的信使核糖核酸前体在HeLa细胞提取物中的剪接
Cell. 1983 Nov;35(1):89-99. doi: 10.1016/0092-8674(83)90211-8.
10
Splicing of messenger RNA precursors is inhibited by antisera to small nuclear ribonucleoprotein.信使核糖核酸前体的剪接受到针对小核核糖核蛋白的抗血清的抑制。
Cell. 1983 Nov;35(1):101-7. doi: 10.1016/0092-8674(83)90212-x.