Title: Structures of the CRISPR genome integration complex

Abstract

CRISPR-Cas systems depend on the Cas1-Cas2 integrase to capture and integrate short foreign DNA fragments into the CRISPR locus, enabling adaptation to new viruses. In this paper, we present crystal structures of Cas1-Cas2 bound to both donor and target DNA in intermediate and product integration complexes, as well as a cryo–electron microscopy structure of the full CRISPR locus integration complex, including the accessory protein IHF (integration host factor). The structures show unexpectedly that indirect sequence recognition dictates integration site selection by favoring deformation of the repeat and the flanking sequences. IHF binding bends the DNA sharply, bringing an upstream recognition motif into contact with Cas1 to increase both the specificity and efficiency of integration. Lastly, these results explain how the Cas1-Cas2 CRISPR integrase recognizes a sequence-dependent DNA structure to ensure site-selective CRISPR array expansion during the initial step of bacterial adaptive immunity.

Authors:

Wright, Addison V.  ^[1];

• Search DOE PAGES for author "Wright, Addison V."
• Search DOE PAGES for ORCID "0000-0002-1018-4312"

Liu, Jun-Jie  ^[2];

• Search DOE PAGES for author "Liu, Jun-Jie"
• Search DOE PAGES for ORCID "0000-0002-8982-107X"

Knott, Gavin J.  ^[1];

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• Search DOE PAGES for ORCID "0000-0002-9007-6273"

Doxzen, Kevin W.  ^[3];

• Search DOE PAGES for author "Doxzen, Kevin W."
• Search DOE PAGES for ORCID "0000-0002-3616-6569"

Nogales, Eva  ^[4];

• Search DOE PAGES for author "Nogales, Eva"
• Search DOE PAGES for ORCID "0000-0001-9816-3681"

Doudna, Jennifer A.  ^[5]

• Search DOE PAGES for author "Doudna, Jennifer A."
• Search DOE PAGES for ORCID "0000-0001-9161-999X"

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+ Show Author Affiliations

1. Univ. of California, Berkeley, CA (United States). Department of Molecular and Cell Biology
2. Univ. of California, Berkeley, CA (United States). Department of Molecular and Cell Biology; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Molecular Biophysics and Integrated Bioimaging Division
3. Univ. of California, Berkeley, CA (United States). Biophysics Graduate Group
4. Univ. of California, Berkeley, CA (United States). Department of Molecular and Cell Biology and Howard Hughes Medical Institute; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Molecular Biophysics and Integrated Bioimaging Division
5. Univ. of California, Berkeley, CA (United States). Department of Molecular and Cell Biology, Biophysics Graduate Group, Howard Hughes Medical Institute, Department of Chemistry, Innovative Genomics Institute, and Center for RNA Systems Biology; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Molecular Biophysics and Integrated Bioimaging Division

Publication Date:

Thu Jul 20 00:00:00 EDT 2017

Research Org.:

Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Org.:

USDOE Office of Science (SC), Basic Energy Sciences (BES); USDOE Office of Science (SC), Biological and Environmental Research (BER)

OSTI Identifier:

1426734

Grant/Contract Number:  

AC02-05CH11231; AC02-76SF00515

Resource Type:

Accepted Manuscript

Journal Name:

Science

Additional Journal Information:

Journal Volume: 357; Journal Issue: 6356; Journal ID: ISSN 0036-8075

Publisher:

AAAS

Country of Publication:

United States

Language:

English

Subject:

59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES