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Title: Cryo-EM Structures Reveal Mechanism and Inhibition of DNA Targeting by a CRISPR-Cas Surveillance Complex

Abstract

Prokaryotic cells possess CRISPR-mediated adaptive immune systems that protect them from foreign genetic elements, such as invading viruses. Here, a central element of this immune system is an RNA-guided surveillance complex capable of targeting non-self DNA or RNA for degradation in a sequence- and site-specific manner analogous to RNA interference. Although the complexes display considerable diversity in their composition and architecture, many basic mechanisms underlying target recognition and cleavage are highly conserved. Using cryoelectron microscopy (cryo-EM), we show that the binding of target double-stranded DNA (dsDNA) to a type I-F CRISPR system yersinia (Csy) surveillance complex leads to large quaternary and tertiary structural changes in the complex that are likely necessary in the pathway leading to target dsDNA degradation by a trans-acting helicase-nuclease. Comparison of the structure of the surveillance complex before and after dsDNA binding, or in complex with three virally encoded anti-CRISPR suppressors that inhibit dsDNA binding, reveals mechanistic details underlying target recognition and inhibition.

Authors:
 [1];  [1];  [2];  [1];  [1];  [1];  [3];  [3];  [4];  [1];  [2];  [1]
  1. National Inst. of Health (NIH), Bethesda, MD (United States)
  2. Memorial Sloan Kettering Cancer Center, New York, NY (United States)
  3. New York Structural Biology Center, New York, NY (United States)
  4. National Inst. of Health (NIH), Bethesda, MD (United States); Frederick National Lab. for Cancer Research, MD (United States)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
National Inst. of Health; Memorial Sloan Kettering Cancer Center; Center for Cancer Research, National Cancer Inst.; Simons Foundation; Agouron Inst.
OSTI Identifier:
1430317
Grant/Contract Number:  
[HHSN261200800001E; P30CA008748; GM104962; 349247; GM103310; F00316; S10 Od019994-01]
Resource Type:
Accepted Manuscript
Journal Name:
Cell
Additional Journal Information:
[ Journal Volume: 171; Journal Issue: 2]; Journal ID: ISSN 0092-8674
Publisher:
Elsevier
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; CRISPR-Cas Csy complexes; CRISPR-Cas systems; adaptive immunity; PAM recognition; DNA-protein interactions; conformational transitions; gene editing; anti-CRISPR inhibitors; cryoelectron microscopy

Citation Formats

Guo, Tai Wei, Bartesaghi, Alberto, Yang, Hui, Falconieri, Veronica, Rao, Prashant, Merk, Alan, Eng, Edward T., Raczkowski, Ashleigh M., Fox, Tara, Earl, Lesley A., Patel, Dinshaw J., and Subramaniam, Sriram. Cryo-EM Structures Reveal Mechanism and Inhibition of DNA Targeting by a CRISPR-Cas Surveillance Complex. United States: N. p., 2017. Web. doi:10.1016/j.cell.2017.09.006.
Guo, Tai Wei, Bartesaghi, Alberto, Yang, Hui, Falconieri, Veronica, Rao, Prashant, Merk, Alan, Eng, Edward T., Raczkowski, Ashleigh M., Fox, Tara, Earl, Lesley A., Patel, Dinshaw J., & Subramaniam, Sriram. Cryo-EM Structures Reveal Mechanism and Inhibition of DNA Targeting by a CRISPR-Cas Surveillance Complex. United States. doi:10.1016/j.cell.2017.09.006.
Guo, Tai Wei, Bartesaghi, Alberto, Yang, Hui, Falconieri, Veronica, Rao, Prashant, Merk, Alan, Eng, Edward T., Raczkowski, Ashleigh M., Fox, Tara, Earl, Lesley A., Patel, Dinshaw J., and Subramaniam, Sriram. Thu . "Cryo-EM Structures Reveal Mechanism and Inhibition of DNA Targeting by a CRISPR-Cas Surveillance Complex". United States. doi:10.1016/j.cell.2017.09.006. https://www.osti.gov/servlets/purl/1430317.
@article{osti_1430317,
title = {Cryo-EM Structures Reveal Mechanism and Inhibition of DNA Targeting by a CRISPR-Cas Surveillance Complex},
author = {Guo, Tai Wei and Bartesaghi, Alberto and Yang, Hui and Falconieri, Veronica and Rao, Prashant and Merk, Alan and Eng, Edward T. and Raczkowski, Ashleigh M. and Fox, Tara and Earl, Lesley A. and Patel, Dinshaw J. and Subramaniam, Sriram},
abstractNote = {Prokaryotic cells possess CRISPR-mediated adaptive immune systems that protect them from foreign genetic elements, such as invading viruses. Here, a central element of this immune system is an RNA-guided surveillance complex capable of targeting non-self DNA or RNA for degradation in a sequence- and site-specific manner analogous to RNA interference. Although the complexes display considerable diversity in their composition and architecture, many basic mechanisms underlying target recognition and cleavage are highly conserved. Using cryoelectron microscopy (cryo-EM), we show that the binding of target double-stranded DNA (dsDNA) to a type I-F CRISPR system yersinia (Csy) surveillance complex leads to large quaternary and tertiary structural changes in the complex that are likely necessary in the pathway leading to target dsDNA degradation by a trans-acting helicase-nuclease. Comparison of the structure of the surveillance complex before and after dsDNA binding, or in complex with three virally encoded anti-CRISPR suppressors that inhibit dsDNA binding, reveals mechanistic details underlying target recognition and inhibition.},
doi = {10.1016/j.cell.2017.09.006},
journal = {Cell},
number = [2],
volume = [171],
place = {United States},
year = {2017},
month = {10}
}

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