The Making of a Slicer: Activation of Human Argonaute-1

Faehnle, Christopher R, Elkayam, Elad, Haase, Astrid D, Hannon, Gregory J, Joshua-Tor, Leemor (2013) The Making of a Slicer: Activation of Human Argonaute-1. Cell Reports, 3 (6). pp. 1901-1909. ISSN 2211-1247

URL: http://www.ncbi.nlm.nih.gov/pubmed/23746446
DOI: 10.1016/j.celrep.2013.05.033

Abstract

Summary Argonautes are the central protein component in small RNA silencing pathways. Of the four human Argonautes (hAgo1–hAgo4) only hAgo2 is an active slicer. We determined the structure of hAgo1 bound to endogenous copurified RNAs to 1.75 Å resolution and hAgo1 loaded with let-7 microRNA to 2.1 Å. Both structures are strikingly similar to the structures of hAgo2. A conserved catalytic tetrad within the PIWI domain of hAgo2 is required for its slicing activity. Completion of the tetrad, combined with a mutation on a loop adjacent to the active site of hAgo1, results in slicer activity that is substantially enhanced by swapping in the N domain of hAgo2. hAgo3, with an intact tetrad, becomes an active slicer by swapping the N domain of hAgo2 without additional mutations. Intriguingly, the elements that make Argonaute an active slicer involve a sophisticated interplay between the active site and more distant regions of the enzyme.

Item Type: Paper
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > argonaute proteins
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types
CSHL Authors:
Communities: CSHL Cancer Center Program > Cancer Genetics
CSHL Cancer Center Program > Gene Regulation and Cell Proliferation
CSHL Cancer Center Shared Resources > DNA Sequencing Service
CSHL Cancer Center Shared Resources > Mass Spectrometry Service
CSHL Cancer Center Shared Resources > Next Generation Sequencing Service
CSHL Cancer Center Shared Resources > Proteomics Service
CSHL labs > Hannon lab
CSHL labs > Joshua-Tor lab
Depositing User: Matt Covey
Date Deposited: 24 Jun 2013 20:33
Last Modified: 01 Sep 2017 16:19
PMCID: PMC3769929
Related URLs:
Dataset ID:
URI: http://repository.cshl.edu/id/eprint/28365

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