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Title: Crystal Structure of a CRISPR RNA-guided Surveillance Complex Bound to a ssDNA Target

Abstract

In prokaryotes, RNA derived from type I and type III CRISPR loci direct large ribonucleoprotein complexes to destroy invading bacteriophage and plasmids. In Escherichia coli, this 405-kilodalton complex is called Cascade. We report the crystal structure of Cascade bound to a single-stranded DNA (ssDNA) target at a resolution of 3.03 angstroms. The structure reveals that the CRISPR RNA and target strands do not form a double helix but instead adopt an underwound ribbon-like structure. This noncanonical structure is facilitated by rotation of every sixth nucleotide out of the RNA-DNA hybrid and is stabilized by the highly interlocked organization of protein subunits. These studies provide insight into both the assembly and the activity of this complex and suggest a mechanism to enforce fidelity of target binding.

Authors:
 [1]; ;  [1]
  1. Johns Hopkins Univ., Baltimore, MD (United States)
Publication Date:
Research Org.:
Brookhaven National Laboratory (BNL), Upton, NY (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1228985
Report Number(s):
BNL-111060-2015-JA
Journal ID: ISSN 1095-9203
DOE Contract Number:  
SC00112704
Resource Type:
Journal Article
Journal Name:
Science (New York, N.Y., Online)
Additional Journal Information:
Journal Volume: 345; Journal Issue: 6203; Journal ID: ISSN 1095-9203
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Mulepati, Sabin, Heroux, Annie, and Bailey, Scott. Crystal Structure of a CRISPR RNA-guided Surveillance Complex Bound to a ssDNA Target. United States: N. p., 2014. Web. doi:10.1126/science.1256996.
Mulepati, Sabin, Heroux, Annie, & Bailey, Scott. Crystal Structure of a CRISPR RNA-guided Surveillance Complex Bound to a ssDNA Target. United States. doi:10.1126/science.1256996.
Mulepati, Sabin, Heroux, Annie, and Bailey, Scott. Fri . "Crystal Structure of a CRISPR RNA-guided Surveillance Complex Bound to a ssDNA Target". United States. doi:10.1126/science.1256996.
@article{osti_1228985,
title = {Crystal Structure of a CRISPR RNA-guided Surveillance Complex Bound to a ssDNA Target},
author = {Mulepati, Sabin and Heroux, Annie and Bailey, Scott},
abstractNote = {In prokaryotes, RNA derived from type I and type III CRISPR loci direct large ribonucleoprotein complexes to destroy invading bacteriophage and plasmids. In Escherichia coli, this 405-kilodalton complex is called Cascade. We report the crystal structure of Cascade bound to a single-stranded DNA (ssDNA) target at a resolution of 3.03 angstroms. The structure reveals that the CRISPR RNA and target strands do not form a double helix but instead adopt an underwound ribbon-like structure. This noncanonical structure is facilitated by rotation of every sixth nucleotide out of the RNA-DNA hybrid and is stabilized by the highly interlocked organization of protein subunits. These studies provide insight into both the assembly and the activity of this complex and suggest a mechanism to enforce fidelity of target binding.},
doi = {10.1126/science.1256996},
journal = {Science (New York, N.Y., Online)},
issn = {1095-9203},
number = 6203,
volume = 345,
place = {United States},
year = {2014},
month = {9}
}