3HO1

Crystal structure of T. thermophilus Argonaute N546 mutant protein complexed with DNA guide strand and 12-nt RNA target strand

3HO1 の概要

• エントリーDOI: 10.2210/pdb3ho1/pdb
• 関連するPDBエントリー: 3DLB 3DLH 3F73 3HGE 3HJF 3HK2
• 分子名称: argonaute, 5'-D(P*TP*GP*AP*GP*GP*TP*AP*GP*TP*AP*GP*GP*TP*TP*GP*TP*AP*T*AP*GP*T)-3', 5'-R(P*CP*CP*UP*AP*CP*UP*AP*CP*CP*UP*CP*G)-3', ... (5 entities in total)
• 機能のキーワード: argonaute, protein-dna-rna complex, nucleic acid binding protein-dna-rna complex, nucleic acid binding protein/dna/rna
• 由来する生物種: Thermus thermophilus
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 87048.57

構造登録者

Wang, Y.,Li, H.,Sheng, G.,Patel, D.J. (登録日: 2009-06-01, 公開日: 2009-10-06, 最終更新日: 2023-09-06)

主引用文献

Wang, Y.,Juranek, S.,Li, H.,Sheng, G.,Wardle, G.S.,Tuschl, T.,Patel, D.J.
Nucleation, propagation and cleavage of target RNAs in Ago silencing complexes.
Nature, 461:754-761, 2009

PubMed Abstract: The slicer activity of the RNA-induced silencing complex resides within its Argonaute (Ago) component, in which the PIWI domain provides the catalytic residues governing guide-strand mediated site-specific cleavage of target RNA. Here we report on structures of ternary complexes of Thermus thermophilus Ago catalytic mutants with 5'-phosphorylated 21-nucleotide guide DNA and complementary target RNAs of 12, 15 and 19 nucleotides in length, which define the molecular basis for Mg(2+)-facilitated site-specific cleavage of the target. We observe pivot-like domain movements within the Ago scaffold on proceeding from nucleation to propagation steps of guide-target duplex formation, with duplex zippering beyond one turn of the helix requiring the release of the 3'-end of the guide from the PAZ pocket. Cleavage assays on targets of various lengths supported this model, and sugar-phosphate-backbone-modified target strands showed the importance of structural and catalytic divalent metal ions observed in the crystal structures.

PubMed: 19812667
DOI: 10.1038/nature08434

実験手法

X-RAY DIFFRACTION (2.6 Å)