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Structure reveals mechanism of CRISPR RNA-guided nuclease recruitment and anti-CRISPR viral mimicry

View ORCID ProfileMaryClare F. Rollins, View ORCID ProfileSaikat Chowdhury, Joshua Carter, Sarah M. Golden, Heini M. Miettinen, Andrew Santiago-Frangos, Dominick Faith, C. Martin Lawrence, View ORCID ProfileGabriel C. Lander, View ORCID ProfileBlake Wiedenheft
doi: https://doi.org/10.1101/453720
MaryClare F. Rollins
1Department of Microbiology and Immunology, Montana State University, Bozeman, MT 59717, USA
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Saikat Chowdhury
3Department of Integrative Structural and Computational Biology, Scripps Research, La Jolla, CA 92037, USA.
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Joshua Carter
1Department of Microbiology and Immunology, Montana State University, Bozeman, MT 59717, USA
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Sarah M. Golden
1Department of Microbiology and Immunology, Montana State University, Bozeman, MT 59717, USA
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Heini M. Miettinen
1Department of Microbiology and Immunology, Montana State University, Bozeman, MT 59717, USA
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Andrew Santiago-Frangos
1Department of Microbiology and Immunology, Montana State University, Bozeman, MT 59717, USA
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Dominick Faith
1Department of Microbiology and Immunology, Montana State University, Bozeman, MT 59717, USA
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C. Martin Lawrence
2Department of Chemistry and Biochemistry, Montana State University, Bozeman, MT 59717, USA
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Gabriel C. Lander
3Department of Integrative Structural and Computational Biology, Scripps Research, La Jolla, CA 92037, USA.
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  • For correspondence: glander@scripps.edu bwiedenheft@gmail.com
Blake Wiedenheft
1Department of Microbiology and Immunology, Montana State University, Bozeman, MT 59717, USA
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  • ORCID record for Blake Wiedenheft
  • For correspondence: glander@scripps.edu bwiedenheft@gmail.com
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Abstract

Bacteria and archaea have evolved sophisticated adaptive immune systems that rely on CRISPR RNA (crRNA)-guided detection and nuclease-mediated elimination of invading nucleic acids. Here we present the cryo-EM structure of the type I-F CRISPR RNA-guided surveillance complex (Csy complex) from Pseudomonas aeruginosa bound to a double-stranded DNA target. Comparison of this structure to previously determined structures of this complex reveals a Ȉ180-degree rotation of the C-terminal helical bundle on the “large” Cas8f subunit. We show that the dsDNA-induced conformational change in Cas8f exposes a Cas2/3 “nuclease recruitment helix” that is structurally homologous to a virally encoded anti-CRISPR protein (AcrIF3). Structural homology between Cas8f and AcrIF3 suggests that AcrIF3 is a mimic of the Cas8f “nuclease recruitment helix”, implying that cas genes may sometimes serve as genetic fodder for the evolution of anti-CRISPRs.

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Posted October 26, 2018.
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Structure reveals mechanism of CRISPR RNA-guided nuclease recruitment and anti-CRISPR viral mimicry
MaryClare F. Rollins, Saikat Chowdhury, Joshua Carter, Sarah M. Golden, Heini M. Miettinen, Andrew Santiago-Frangos, Dominick Faith, C. Martin Lawrence, Gabriel C. Lander, Blake Wiedenheft
bioRxiv 453720; doi: https://doi.org/10.1101/453720
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Structure reveals mechanism of CRISPR RNA-guided nuclease recruitment and anti-CRISPR viral mimicry
MaryClare F. Rollins, Saikat Chowdhury, Joshua Carter, Sarah M. Golden, Heini M. Miettinen, Andrew Santiago-Frangos, Dominick Faith, C. Martin Lawrence, Gabriel C. Lander, Blake Wiedenheft
bioRxiv 453720; doi: https://doi.org/10.1101/453720

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