1Z25

Structure of P.furiosus Argonaute with bound Mn2+

Summary for 1Z25

• Entry DOI: 10.2210/pdb1z25/pdb
• Related: 1U04 1Z26
• Descriptor: Argonaute, MANGANESE (II) ION (3 entities in total)
• Functional Keywords: argonaute, piwi, active site, rnaseh, mg2+, rnai, gene regulation
• Biological source: Pyrococcus furiosus
• Total number of polymer chains: 1
• Total formula weight: 90652.11

Authors

Rivas, F.V.,Tolia, N.H.,Song, J.J.,Aragon, J.P.,Liu, J.,Hannon, G.J.,Joshua-Tor, L. (deposition date: 2005-03-07, release date: 2005-04-05, Last modification date: 2024-02-14)

Primary citation

Rivas, F.V.,Tolia, N.H.,Song, J.J.,Aragon, J.P.,Liu, J.,Hannon, G.J.,Joshua-Tor, L.
Purified Argonaute2 and an siRNA form recombinant human RISC.
Nat.Struct.Mol.Biol., 12:340-349, 2005

PubMed Abstract: Genetic, biochemical and structural studies have implicated Argonaute proteins as the catalytic core of the RNAi effector complex, RISC. Here we show that recombinant, human Argonaute2 can combine with a small interfering RNA (siRNA) to form minimal RISC that accurately cleaves substrate RNAs. Recombinant RISC shows many of the properties of RISC purified from human or Drosophila melanogaster cells but also has surprising features. It shows no stimulation by ATP, suggesting that factors promoting product release are missing from the recombinant enzyme. The active site is made up of a unique Asp-Asp-His (DDH) motif. In the RISC reconstitution system, the siRNA 5' phosphate is important for the stability and the fidelity of the complex but is not essential for the creation of an active enzyme. These studies demonstrate that Argonaute proteins catalyze mRNA cleavage within RISC and provide a source of recombinant enzyme for detailed biochemical studies of the RNAi effector complex.

PubMed: 15800637
DOI: 10.1038/nsmb918

Experimental method

X-RAY DIFFRACTION (2.7 Å)