Structural features of Argonaute–GW182 protein interactions
Autor: | Dierk Niessing, Michael Sattler, Franz Herzog, Ruedi Aebersold, Gunter Meister, Janina Pfaff, Janosch Hennig |
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Jazyk: | angličtina |
Rok vydání: | 2013 |
Předmět: |
Models
Molecular Magnetic Resonance Spectroscopy Protein family Protein Conformation Genetic Vectors Fluorescence Polarization RNA-binding protein Plasma protein binding Biology Autoantigens Mass Spectrometry Protein–protein interaction Protein structure miRNA Ago2 GW-protein TNRC6Am TNRC6B TNRC6C structural biology NMR Gene regulation smnall RNA-mediated gene silencing RNAi RNA interference Humans Immunoprecipitation Gene silencing Genetics Multidisciplinary Circular Dichroism RNA-Binding Proteins Argonaute Cell biology HEK293 Cells PNAS Plus Gene Expression Regulation Multiprotein Complexes Argonaute Proteins RNA Interference Baculoviridae Protein Binding |
Zdroj: | Proc. Natl. Acad. Sci. U.S.A. 110, 3770-3779 (2013) |
Popis: | MicroRNAs (miRNAs) guide Argonaute (Ago) proteins to target mRNAs, leading to gene silencing. However, Ago proteins are not the actual mediators of gene silencing but interact with a member of the GW182 protein family (also known as GW proteins), which coordinates all downstream steps in gene silencing. GW proteins contain an N-terminal Ago-binding domain that is characterized by multiple GW repeats and a C-terminal silencing domain with several globular domains. Within the Ago-binding domain, Trp residues mediate the direct interaction with the Ago protein. Here, we have characterized the interaction of Ago proteins with GW proteins in molecular detail. Using biochemical and NMR experiments, we show that only a subset of Trp residues engage in Ago interactions. The Trp residues are located in intrinsically disordered regions, where flanking residues mediate additional weak interactions, that might explain the importance of specific tryptophans. Using cross-linking followed by mass spectrometry, we map the GW protein interactions with Ago2, which allows for structural modeling of Ago-GW182 interaction. Our data further indicate that the Ago-GW protein interaction might be a two-step process involving the sequential binding of two tryptophans separated by a spacer with a minimal length of 10 aa. |
Databáze: | OpenAIRE |
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