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连接I组核酶的两个结构域以形成催化核心。

Joining the two domains of a group I ribozyme to form the catalytic core.

作者信息

Tanner M A, Cech T R

机构信息

Department of Chemistry and Biochemistry, Howard Hughes Medical Institute, University of Colorado, Boulder, Colorado 80309-0215, USA.

出版信息

Science. 1997 Feb 7;275(5301):847-9. doi: 10.1126/science.275.5301.847.

Abstract

Self-splicing group I introns, like other large catalytic RNAs, contain structural domains. Although the crystal structure of one of these domains has been determined by x-ray analysis, its connection to the other major domain that contains the guanosine-binding site has not been known. Site-directed mutagenesis and kinetic analysis of RNA splicing were used to identify a base triple in the conserved core of both a cyanobacterial (Anabaena) and a eukaryotic (Tetrahymena) group I intron. This long-range interaction connects a sequence adjacent to the guanosine-binding site with the domain implicated in coordinating the 5' splice site helix, and it thereby contributes to formation of the active site. The resulting five-strand junction, in which a short helix forms base triples with three separate strands in the Tetrahymena intron, reveals exceptionally dense packing of RNA.

摘要

自我剪接的I组内含子与其他大型催化RNA一样,都含有结构域。尽管其中一个结构域的晶体结构已通过X射线分析确定,但其与包含鸟苷结合位点的另一个主要结构域的连接方式尚不清楚。通过对RNA剪接进行定点诱变和动力学分析,在蓝细菌(鱼腥藻)和真核生物(嗜热四膜虫)的I组内含子的保守核心区域中鉴定出一个碱基三联体。这种长程相互作用将与鸟苷结合位点相邻的序列与参与协调5'剪接位点螺旋的结构域连接起来,从而有助于活性位点的形成。由此形成的五链连接结构中,嗜热四膜虫内含子中的一个短螺旋与三条独立的链形成碱基三联体,显示出RNA异常紧密的堆积。

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