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Title: The Inside-Out Mechanism of Dicers from Budding Yeasts

Abstract

The Dicer ribonuclease III (RNase III) enzymes process long double-stranded RNA (dsRNA) into small interfering RNAs (siRNAs) that direct RNA interference. Here, we describe the structure and activity of a catalytically active fragment of Kluyveromyces polysporus Dcr1, which represents the noncanonical Dicers found in budding yeasts. The crystal structure revealed a homodimer resembling that of bacterial RNase III but extended by a unique N-terminal domain, and it identified additional catalytic residues conserved throughout eukaryotic RNase III enzymes. Biochemical analyses showed that Dcr1 dimers bind cooperatively along the dsRNA substrate such that the distance between consecutive active sites determines the length of the siRNA products. Thus, unlike canonical Dicers, which successively remove siRNA duplexes from the dsRNA termini, budding-yeast Dicers initiate processing in the interior and work outward. The distinct mechanism of budding-yeast Dicers establishes a paradigm for natural molecular rulers and imparts substrate preferences with ramifications for biological function.

Authors:
; ; ;  [1];  [2]
  1. (Whitehead)
  2. (
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
FOREIGNNSFOTHERNIH
OSTI Identifier:
1024047
Resource Type:
Journal Article
Journal Name:
Cell
Additional Journal Information:
Journal Volume: 146; Journal Issue: (2) ; 07, 2011; Journal ID: ISSN 0092-8674
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; BIOLOGICAL FUNCTIONS; CRYSTAL STRUCTURE; DIMERS; ENZYMES; PROCESSING; RESIDUES; RNA; RNA-ASE; SUBSTRATES; YEASTS

Citation Formats

Weinberg, David E., Nakanishi, Kotaro, Patel, Dinshaw J., Bartel, David P., and MSKCC). The Inside-Out Mechanism of Dicers from Budding Yeasts. United States: N. p., 2011. Web. doi:10.1016/j.cell.2011.06.021.
Weinberg, David E., Nakanishi, Kotaro, Patel, Dinshaw J., Bartel, David P., & MSKCC). The Inside-Out Mechanism of Dicers from Budding Yeasts. United States. doi:10.1016/j.cell.2011.06.021.
Weinberg, David E., Nakanishi, Kotaro, Patel, Dinshaw J., Bartel, David P., and MSKCC). Tue . "The Inside-Out Mechanism of Dicers from Budding Yeasts". United States. doi:10.1016/j.cell.2011.06.021.
@article{osti_1024047,
title = {The Inside-Out Mechanism of Dicers from Budding Yeasts},
author = {Weinberg, David E. and Nakanishi, Kotaro and Patel, Dinshaw J. and Bartel, David P. and MSKCC)},
abstractNote = {The Dicer ribonuclease III (RNase III) enzymes process long double-stranded RNA (dsRNA) into small interfering RNAs (siRNAs) that direct RNA interference. Here, we describe the structure and activity of a catalytically active fragment of Kluyveromyces polysporus Dcr1, which represents the noncanonical Dicers found in budding yeasts. The crystal structure revealed a homodimer resembling that of bacterial RNase III but extended by a unique N-terminal domain, and it identified additional catalytic residues conserved throughout eukaryotic RNase III enzymes. Biochemical analyses showed that Dcr1 dimers bind cooperatively along the dsRNA substrate such that the distance between consecutive active sites determines the length of the siRNA products. Thus, unlike canonical Dicers, which successively remove siRNA duplexes from the dsRNA termini, budding-yeast Dicers initiate processing in the interior and work outward. The distinct mechanism of budding-yeast Dicers establishes a paradigm for natural molecular rulers and imparts substrate preferences with ramifications for biological function.},
doi = {10.1016/j.cell.2011.06.021},
journal = {Cell},
issn = {0092-8674},
number = (2) ; 07, 2011,
volume = 146,
place = {United States},
year = {2011},
month = {9}
}