2M58

Structure of 2'-5' AG1 lariat forming ribozyme in its inactive state

2M58 の概要

• エントリーDOI: 10.2210/pdb2m58/pdb
• NMR情報: BMRB: 19040
• 分子名称: RNA (59-MER) (1 entity in total)
• 機能のキーワード: rna, lariat, 2'-5' branching
• 由来する生物種: synthetic
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 19163.36

構造登録者

Carlomagno, T.,Amata, I.,Codutti, L.,Falb, M.,Fohrer, J.,Simon, B. (登録日: 2013-02-18, 公開日: 2013-04-03, 最終更新日: 2024-05-15)

主引用文献

Carlomagno, T.,Amata, I.,Codutti, L.,Falb, M.,Fohrer, J.,Masiewicz, P.,Simon, B.
Structural principles of RNA catalysis in a 2'-5' lariat-forming ribozyme.
J.Am.Chem.Soc., 135:4403-4411, 2013

PubMed Abstract: RNA-catalyzed lariat formation is present in both eukaryotes and prokaryotes. To date we lack structural insights into the catalytic mechanism of lariat-forming ribozymes. Here, we study an artificial 2'-5' AG1 lariat-forming ribozyme that shares the sequence specificity of lariat formation with the pre-mRNA splicing reaction. Using NMR, we solve the structure of the inactive state of the ribozyme in the absence of magnesium. The reaction center 5'-guanosine appears to be part of a helix with an exceptionally widened major groove, while the lariat-forming A48 is looped out at the apex of a pseudoknot. The model of the active state built by mutational analysis, molecular modeling, and small-angle X-ray scattering suggests that A48 is recognized by a conserved adenosine, juxtaposed to the 5'-guanosine in one base-pair step distance, while the G1-N7 coordinates a magnesium ion essential for the activation of the nucleophile. Our findings offer implications for lariat formation in RNA enzymes including the mechanism of the recognition of the branch-site adenosine.

PubMed: 23472843
DOI: 10.1021/ja311868t

実験手法

SOLUTION NMR