Phage AcrIIA2 DNA Mimicry: Structural Basis of the CRISPR and Anti-CRISPR Arms Race

Liu, Liang; Yin, Maolu; Wang, Min; Wang, Yanli

doi:10.1016/j.molcel.2018.11.011

Cited by 87 publications

(72 citation statements)

References 37 publications

(57 reference statements)

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“…Former mechanistic characterizations of Acr proteins have revealed a remarkable diversity of inhibitory functions (Davidson et al, 2020). Interestingly, relatively low MWs and pIs have been reported for Acrs inhibiting CRISPR-Cas systems via DNA mimicry (Jiang et al, 2019;Liu et al, 2019), suggesting a putative link between low MW/pI values and mechanistic inhibitory function. Given the small size and negative charges of AcrIF18* and AcrIF15 (Supplementary Figure S2), and their Cascade DNA binding inhibitory activities (Supplementary Figure S3), it is possible that they function as DNA mimics -analogous to AcrIF2 (Chowdhury et al, 2017), AcrIF10 (Guo et al, 2017), AcrIIA2 (Shin et al, 2017;Liu et al, 2019) and AcrIIA4 (Dong et al, 2017).…”

Section: Discussionmentioning

confidence: 99%

Discovery of multiple anti-CRISPRs uncovers anti-defense gene clustering in mobile genetic elements

Pinilla‐Redondo

Shehreen

Marino

et al. 2020

Preprint

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Many prokaryotes employ CRISPR-Cas systems to combat invading mobile genetic elements (MGEs). In response, some MGEs have evolved Anti-CRISPR (Acr) proteins to bypass this immunity, yet the diversity, distribution and spectrum of activity of this immune evasion strategy remain largely unknown. Here, we uncover 11 new type I anti-CRISPR genes encoded on numerous chromosomal and extrachromosomal mobile genetic elements within Enterobacteriaceae and Pseudomonas. Candidate genes were identified adjacent to anti-CRISPR associated gene 5 (aca5) and assayed against a panel of six type I systems: I-F (Pseudomonas, Pectobacterium, and Serratia), I-E (Pseudomonas and Serratia), and I-C (Pseudomonas), revealing the type I-F and/or I-E acr genes and a new aca (aca9). We find that acr genes not only associate with other acr genes, but also with inhibitors of distinct bacterial defense systems. These genomic regions appear to be "anti-defense islands", reminiscent of the clustered arrangement of "defense islands" in prokaryotic genomes. Our findings expand on the diversity of CRISPR-Cas inhibitors and reveal the potential exploitation of acr loci neighborhoods for identifying new anti-defense systems.

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Section: Discussionmentioning

confidence: 99%

Discovery of multiple anti-CRISPRs uncovers anti-defense gene clustering in mobile genetic elements

Pinilla‐Redondo

Shehreen

Marino

et al. 2020

Preprint

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“…Combining electrophoretic mobility shift assays (EMASs), fluorescence polarization assays and image assays, J. Lee et al showed that both AcrIIC4 and AcrIIC5 prevent NmeCas9 from binding to DNA while having no effect on sgRNA loading . Recently, two structural studies revealed that AcrIIA2 acts as a DNA mimic, blocking the PAM recognition residues and subsequently preventing dsDNA recognition and binding, which is similar to the actions of AcrIIA4 …”

Section: Acr Mechanismsmentioning

confidence: 99%

Anti‐CRISPRs: The natural inhibitors for CRISPR‐Cas systems

Zhang

Song

Tian

2019

Anim Models and Exp Med

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CRISPR (clustered regularly interspaced short palindromic repeats)‐Cas (CRISPR associated protein) systems serve as the adaptive immune system by which prokaryotes defend themselves against phages. It has also been developed into a series of powerful gene‐editing tools. As the natural inhibitors of CRISPR‐Cas systems, anti‐CRISPRs (Acrs) can be used as the “off‐switch” for CRISPR‐Cas systems to limit the off‐target effects caused by Cas9. Since the discovery of CRISPR‐Cas systems, much research has focused on the identification, mechanisms and applications of Acrs. In light of the rapid development and scientific significance of this field, this review summarizes the history and research status of Acrs, and considers future applications.

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“…AcrIIA1 acts through a distinct structural and functional characteristic, forming a dimer with a two helical-domain structure and functions via nucleic acid recognition, suggesting that AcrIIA1 functions through a unique mechanism that is not yet clear [78]. A recent approach provided a more detailed mechanism of the AcrIIA2-mediated inhibition of spCas9, showing that AcrIIA2 as a DNA mimic interacts with the PAM recognition residues of Cas9, thereby preventing target dsDNA detection [79]. Basgall et al [80] demonstrated that AcrIIA2 and AcrIIA4 are able to inhibit the functional activity of Cas9 in vivo within haploid yeast and in an active gene-driven system, providing the first documented use of AcrIIA2 and AcrIIA4 to inhibit such a system.…”

Section: Type II Anti-crispr Proteinsmentioning

confidence: 99%

Anti‐CRISPR proteins targeting the CRISPR‐Cas system enrich the toolkit for genetic engineering

2019

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Clustered regularly interspaced short palindromic repeats (CRISPR)‐Cas adaptive immune defense systems, which are widely distributed in bacteria and Archaea, can provide sequence‐specific protection against foreign DNA or RNA in some cases. However, the evolution of defense systems in bacterial hosts did not lead to the elimination of phages, and some phages carry anti‐CRISPR genes that encode products that bind to the components mediating the defense mechanism and thus antagonize CRISPR‐Cas immune systems of bacteria. Given the extensive application of CRISPR‐Cas9 technologies in gene editing, in this review, we focus on the anti‐CRISPR proteins (Acrs) that inhibit CRISPR‐Cas systems for gene editing. We describe the discovery of Acrs in immune systems involving type I, II, and V CRISPR‐Cas immunity, discuss the potential function of Acrs in inactivating type II and V CRISPR‐Cas systems for gene editing and gene modulation, and provide an outlook on the development of important biotechnology tools for genetic engineering using Acrs.

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Phage AcrIIA2 DNA Mimicry: Structural Basis of the CRISPR and Anti-CRISPR Arms Race

Abstract: Highlights d Crystal structure of SpyCas9 in complex with sgRNA and suppressor AcrIIA2 d AcrIIA2 recognizes sgRNA-bound SpyCas9 d AcrIIA2 prevents the target DNA search by blocking the PAM recognition residues d AcrIIA2 disables bacterial Cas9 by competing with target dsDNA binding

Cited by 87 publications

References 37 publications

Discovery of multiple anti-CRISPRs uncovers anti-defense gene clustering in mobile genetic elements

Discovery of multiple anti-CRISPRs uncovers anti-defense gene clustering in mobile genetic elements

Anti‐CRISPRs: The natural inhibitors for CRISPR‐Cas systems

Anti‐CRISPR proteins targeting the CRISPR‐Cas system enrich the toolkit for genetic engineering

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