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Pfaff, J. ; Hennig, J. ; Herzog, F.* ; Aebersold, R.* ; Sattler, M. ; Niessing, D. ; Meister, G.*

Structural features of Argonaute–GW182 protein interactions.

Proc. Natl. Acad. Sci. U.S.A. 110, 3770-3779 (2013)
DOI PMC
Open Access Gold as soon as Publ. Version/Full Text is submitted to ZB.
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.
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Publication type Article: Journal article
Document type Scientific Article
Keywords miRNA; Ago2; GW-protein; TNRC6Am; TNRC6B; TNRC6C; structural biology; NMR; Gene regulation; smnall RNA-mediated gene silencing; RNAi; RNA interference; Multiple Sequence Alignment ; Nmr Chemical-shifts ; Translational Repression ; Mass-spectrometry ; Rna Interference ; Gw182 ; Domain ; Micrornas ; Binding ; Relaxation
ISSN (print) / ISBN 0027-8424
e-ISSN 1091-6490
Journal Proceedings of the National Academy of Sciences of the United States of America
Quellenangaben Volume: 110, Issue: 40, Pages: 3770-3779 Article Number: , Supplement: ,
Publisher National Academy of Sciences
Reviewing status Peer reviewed
Institute(s) Institute of Structural Biology (STB)