Details

Autor(en) / Beteiligte

• Salomon, William E.
• Jolly, Samson M.
• Moore, Melissa J.
• Zamore, Phillip D.
• Serebrov, Victor

Titel

Single-Molecule Imaging Reveals that Argonaute Reshapes the Binding Properties of Its Nucleic Acid Guides

Ist Teil von

• Cell, 2015-07, Vol.162 (1), p.84-95

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2015

Link zum Volltext

Quelle

Free E-Journal (出版社公開部分のみ）

Beschreibungen/Notizen

• Argonaute proteins repress gene expression and defend against foreign nucleic acids using short RNAs or DNAs to specify the correct target RNA or DNA sequence. We have developed single-molecule methods to analyze target binding and cleavage mediated by the Argonaute:guide complex, RISC. We find that both eukaryotic and prokaryotic Argonaute proteins reshape the fundamental properties of RNA:RNA, RNA:DNA, and DNA:DNA hybridization—a small RNA or DNA bound to Argonaute as a guide no longer follows the well-established rules by which oligonucleotides find, bind, and dissociate from complementary nucleic acid sequences. Argonautes distinguish substrates from targets with similar complementarity. Mouse AGO2, for example, binds tighter to miRNA targets than its RNAi cleavage product, even though the cleaved product contains more base pairs. By re-writing the rules for nucleic acid hybridization, Argonautes allow oligonucleotides to serve as specificity determinants with thermodynamic and kinetic properties more typical of RNA-binding proteins than of RNA or DNA. [Display omitted] •Argonaute changes the rate of target finding by pre-organizing a region of its guide•Seed pairing is required for rapid target finding and stable binding•AGO2 releases its cleaved products by destabilizing their interaction with the guide•RISC makes its guide behave more like an RNA-binding protein and less like free RNA Argonaute proteins reshape how oligonucleotides find, bind, and dissociate from complementary nucleic acid sequences. By re-writing the rules, Argonautes allow oligonucleotides to serve as specificity determinants with thermodynamic and kinetic properties more typical of RNA-binding proteins.