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Sequence-specific affinity selection of mammalian splicing complexes.

作者信息

Ryder U, Sproat B S, Lamond A I

机构信息

European Molecular Biology Laboratory, Heidelberg, FRG.

出版信息

Nucleic Acids Res. 1990 Dec 25;18(24):7373-9. doi: 10.1093/nar/18.24.7373.

DOI:10.1093/nar/18.24.7373
PMID:2147994
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC332875/
Abstract

Antisense oligonucleotides made of 2'-OMe RNA are shown to bind specifically and efficiently to targeted sites on pre-mRNA substrates, allowing affinity selection of splicing complexes using streptavidin/biotin chromatography. The position of probe binding to the pre-mRNA influences which type of splicing complex can be selected. The accessibility of pre-mRNA sequences to antisense probes changes during the course of the splicing reaction. U1, U2, U4, U5 and U6 snRNAs are all detected in affinity-selected mammalian splicing complexes. However, antisense oligonucleotides targeted to snRNAs can block the binding of specific snRNPs to pre-mRNA. Quantitative affinity selection analyses show that only a small fraction of snRNPs in a HeLa nuclear splicing extract participate in spliceosome formation.

摘要

相似文献

1
Sequence-specific affinity selection of mammalian splicing complexes.
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2
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3
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4
A block in mammalian splicing occurring after formation of large complexes containing U1, U2, U4, U5, and U6 small nuclear ribonucleoproteins.在包含U1、U2、U4、U5和U6小核核糖核蛋白的大复合体形成后发生的哺乳动物剪接阻断。
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5
U1-U2 snRNPs interaction induced by an RNA complementary to the 5' end sequence of U1 snRNA.由与U1 snRNA 5'端序列互补的RNA诱导的U1-U2 snRNP相互作用。
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6
Antisense probes containing 2-aminoadenosine allow efficient depletion of U5 snRNP from HeLa splicing extracts.含有2-氨基腺苷的反义探针能够有效地从HeLa剪接提取物中耗尽U5 snRNP。
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7
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Nature. 1991 Aug 29;352(6338):818-21. doi: 10.1038/352818a0.
8
Targeted snRNP depletion reveals an additional role for mammalian U1 snRNP in spliceosome assembly.靶向性snRNP缺失揭示了哺乳动物U1 snRNP在剪接体组装中的额外作用。
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