分支点腺苷、协调环和 3' 外显子结合位点对 Sinorhizobium meliloti 组 II 内含子 RmInt1 体内切除的相关性。

Relevance of the branch point adenosine, coordination loop, and 3' exon binding site for in vivo excision of the Sinorhizobium meliloti group II intron RmInt1.

机构信息

Grupo de Ecología Genética, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Calle Profesor Albareda 1, 18008 Granada, Spain.

出版信息

J Biol Chem. 2011 Jun 17;286(24):21154-63. doi: 10.1074/jbc.M110.210013. Epub 2011 Apr 26.

Abstract

Excision of the bacterial group II intron RmInt1 has been demonstrated in vivo, resulting in the formation of both intron lariat and putative intron RNA circles. We show here that the bulged adenosine in domain VI of RmInt1 is required for splicing via the branching pathway, but branch site mutants produce small numbers of RNA molecules in which the first G residue of the intron is linked to the last C residue. Mutations in the coordination loop in domain I reduced splicing efficiency, but branched templates clearly predominated among splicing products. We also found that a single substitution at the EBS3 position (G329C), preventing EBS3-IBS3 pairing, resulted in the production of 50 to 100 times more RNA molecules in which the 5' and 3' extremities were joined. We provide evidence that these intron molecules may correspond to both, intron circles linked by a 2'-5' phosphodiester bond, and tandem, head-to-tail intron copies.

摘要

细菌 Group II 内含子 RmInt1 的切除已在体内得到证实,导致内含子套索和推定内含子 RNA 环的形成。我们在这里表明,RmInt1 结构域 VI 中的凸起腺苷对于通过分支途径进行剪接是必需的,但分支位点突变体产生数量较少的 RNA 分子,其中内含子的第一个 G 残基与最后一个 C 残基相连。结构域 I 中的协调环突变降低了剪接效率,但分支模板显然在剪接产物中占主导地位。我们还发现,EBS3 位置(G329C)的单个取代阻止了 EBS3-IBS3 配对,导致连接 5'和 3'末端的 RNA 分子的产量增加了 50 至 100 倍。我们提供的证据表明,这些内含子分子可能对应于通过 2'-5'磷酸二酯键连接的内含子环,以及串联的、头对头的内含子副本。

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