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低丰度的U11和U12小核核糖核蛋白(snRNP)相互作用形成一个双snRNP复合体。

The low-abundance U11 and U12 small nuclear ribonucleoproteins (snRNPs) interact to form a two-snRNP complex.

作者信息

Wassarman K M, Steitz J A

机构信息

Department of Molecular Biophysics and Biochemistry, Howard Hughes Medical Institute, Yale University, New Haven, Connecticut 06536-0812.

出版信息

Mol Cell Biol. 1992 Mar;12(3):1276-85. doi: 10.1128/mcb.12.3.1276-1285.1992.

Abstract

A novel small nuclear ribonucleoprotein (snRNP) complex containing both U11 and U12 RNAs has been identified in HeLa cell extracts. This U11/U12 snRNP complex can be visualized on glycerol gradients, on native polyacrylamide gels, and by selection with antisense 2'-O-methyl oligoribonucleotides. RNase H-mediated degradation of the U12 snRNA confirmed a direct interaction between the U11 and U12 snRNPs. This snRNP complex is the first to be identified involving low-abundance snRNPs. Selection of the U11/U12 snRNP complex is sensitive to high salt, suggestive of a protein-mediated interaction. Secondary structure analyses revealed several regions of the U11 snRNP accessible for interaction with other RNAs or proteins but no detectable difference between the accessibility of these regions in the U11 monoparticle compared with the U11/U12 snRNP complex. There are also several accessible single-stranded regions in the U12 snRNP, and oligonucleotide-directed RNase H digestion identified nucleotides 28 to 36 of U12 as containing sequences required for the U11/U12 interaction. Both the U12 snRNP and the U11/U12 snRNP complex can be disrupted without altering the cleavage/polyadenylation activity of a nuclear extract.

摘要

在HeLa细胞提取物中发现了一种包含U11和U12 RNA的新型小核核糖核蛋白(snRNP)复合体。这种U11/U12 snRNP复合体可以在甘油梯度、天然聚丙烯酰胺凝胶上观察到,也可以通过用反义2'-O-甲基寡核糖核苷酸进行筛选来观察。核糖核酸酶H介导的U12 snRNA降解证实了U11和U12 snRNP之间的直接相互作用。这种snRNP复合体是首个被鉴定出涉及低丰度snRNP的复合体。U11/U12 snRNP复合体的筛选对高盐敏感,提示存在蛋白质介导的相互作用。二级结构分析揭示了U11 snRNP中几个可与其他RNA或蛋白质相互作用的区域,但与U11单颗粒相比,U11/U12 snRNP复合体中这些区域的可及性没有可检测到的差异。U12 snRNP中也有几个可及的单链区域,寡核苷酸导向的核糖核酸酶H消化确定U12的28至36位核苷酸包含U11/U12相互作用所需的序列。U12 snRNP和U11/U12 snRNP复合体均可被破坏,而不会改变核提取物的切割/聚腺苷酸化活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b862/369560/d474340cb8c0/molcellb00167-0384-a.jpg

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