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Inhibition Mechanism of an Anti-CRISPR Suppressor AcrIIA4 Targeting SpyCas9.

Yang H, Patel DJ
Mol Cell 67 117-127.e5 (2017)
Cited: 82 times
EuropePMC logo PMID: 28602637

Abstract

Prokaryotic CRISPR-Cas adaptive immune systems utilize sequence-specific RNA-guided endonucleases to defend against infection by viruses, bacteriophages, and mobile elements, while these foreign genetic elements evolve diverse anti-CRISPR proteins to overcome the CRISPR-Cas-mediated defense of the host. Recently, AcrIIA2 and AcrIIA4, encoded by Listeria monocytogene prophages, were shown to block the endonuclease activity of type II-A Streptococcus pyogene Cas9 (SpyCas9). We now report the crystal structure of AcrIIA4 in complex with single-guide RNA-bound SpyCas9, thereby establishing that AcrIIA4 preferentially targets critical residues essential for PAM duplex recognition, as well as blocks target DNA access to key catalytic residues lining the RuvC pocket. These structural insights, validated by biochemical assays on key mutants, demonstrate that AcrIIA4 competitively occupies both PAM-interacting and non-target DNA strand cleavage catalytic pockets. Our studies provide insights into anti-CRISPR-mediated suppression mechanisms for inactivating SpyCas9, thereby broadening the applicability of CRISPR-Cas regulatory tools for genome editing.

Reviews - 5vw1 mentioned but not cited (5)

  1. Structures and Strategies of Anti-CRISPR-Mediated Immune Suppression. Wiegand T, Karambelkar S, Bondy-Denomy J, Wiedenheft B. Annu Rev Microbiol 74 21-37 (2020)
  2. Keeping crispr in check: diverse mechanisms of phage-encoded anti-crisprs. Trasanidou D, Gerós AS, Mohanraju P, Nieuwenweg AC, Nobrega FL, Staals RHJ. FEMS Microbiol Lett 366 fnz098 (2019)
  3. Structural insights into the inactivation of CRISPR-Cas systems by diverse anti-CRISPR proteins. Zhu Y, Zhang F, Huang Z. BMC Biol 16 32 (2018)
  4. Insights into the Mechanism of CRISPR/Cas9-Based Genome Editing from Molecular Dynamics Simulations. Bhattacharya S, Satpati P. ACS Omega 8 1817-1837 (2023)
  5. The Many (Inter)faces of Anti-CRISPRs: Modulation of CRISPR-Cas Structure and Dynamics by Mechanistically Diverse Inhibitors. Belato HB, Lisi GP. Biomolecules 13 264 (2023)

Articles - 5vw1 mentioned but not cited (6)



Reviews citing this publication (23)

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  2. Phage-Encoded Anti-CRISPR Defenses. Stanley SY, Maxwell KL. Annu Rev Genet 52 445-464 (2018)
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  5. Applications of CRISPR-Cas Enzymes in Cancer Therapeutics and Detection. Huang CH, Lee KC, Doudna JA. Trends Cancer 4 499-512 (2018)
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  21. A comprehensive appraisal of mechanism of anti-CRISPR proteins: an advanced genome editor to amend the CRISPR gene editing. Choudhary N, Tandi D, Verma RK, Yadav VK, Dhingra N, Ghosh T, Choudhary M, Gaur RK, Abdellatif MH, Gacem A, Eltayeb LB, Alqahtani MS, Yadav KK, Jeon BH. Front Plant Sci 14 1164461 (2023)
  22. Recent advances in the use of CRISPR/Cas for understanding the early development of molecular gaps in glial cells. Barragán-Álvarez CP, Flores-Fernandez JM, Hernández-Pérez OR, Ávila-Gónzalez D, Díaz NF, Padilla-Camberos E, Dublan-García O, Gómez-Oliván LM, Diaz-Martinez NE. Front Cell Dev Biol 10 947769 (2022)
  23. Small Molecules for Enhancing the Precision and Safety of Genome Editing. Shin S, Jang S, Lim D. Molecules 27 6266 (2022)

Articles citing this publication (48)



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