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深入了解 AcrIF14 在抑制 I 型-F CRISPR-Cas 监测复合物中的双重功能。

Insights into the dual functions of AcrIF14 during the inhibition of type I-F CRISPR-Cas surveillance complex.

机构信息

Beijing Advanced Innovation Center for Soft Matter Science and Engineering, Beijing Key Laboratory of Bioprocess, State Key Laboratory of Chemical Resource Engineering, College of Life Science and Technology, Beijing University of Chemical Technology, 100029 Beijing, China.

Ministry of Education Key Laboratory of Protein Science, Beijing Advanced Innovation Center for Structural Biology, Beijing Frontier Research Center for Biological Structure, School of Life Sciences, Tsinghua University, Tsinghua-Peking Center for Life Sciences, 100084 Beijing, China.

出版信息

Nucleic Acids Res. 2021 Sep 27;49(17):10178-10191. doi: 10.1093/nar/gkab738.

DOI:10.1093/nar/gkab738
PMID:34432044
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8464039/
Abstract

CRISPR-Cas systems are bacterial adaptive immune systems, and phages counteract these systems using many approaches such as producing anti-CRISPR (Acr) proteins. Here, we report the structures of both AcrIF14 and its complex with the crRNA-guided surveillance (Csy) complex. Our study demonstrates that apart from interacting with the Csy complex to block the hybridization of target DNA to the crRNA, AcrIF14 also endows the Csy complex with the ability to interact with non-sequence-specific dsDNA as AcrIF9 does. Further structural studies of the Csy-AcrIF14-dsDNA complex and biochemical studies uncover that the PAM recognition loop of the Cas8f subunit of the Csy complex and electropositive patches within the N-terminal domain of AcrIF14 are essential for the non-sequence-specific dsDNA binding to the Csy-AcrIF14 complex, which is different from the mechanism of AcrIF9. Our findings highlight the prevalence of Acr-induced non-specific DNA binding and shed light on future studies into the mechanisms of such Acr proteins.

摘要

CRISPR-Cas 系统是细菌适应性免疫系统，噬菌体通过多种方法来对抗这些系统，例如产生抗 CRISPR（Acr）蛋白。在这里，我们报告了 AcrIF14 及其与 crRNA 引导的监视（Csy）复合物复合物的结构。我们的研究表明，AcrIF14 除了与 Csy 复合物相互作用以阻止靶 DNA 与 crRNA 的杂交外，还赋予 Csy 复合物与非序列特异性 dsDNA 相互作用的能力，就像 AcrIF9 一样。对 Csy-AcrIF14-dsDNA 复合物的进一步结构研究和生化研究表明，Csy 复合物的 Cas8f 亚基的 PAM 识别环和 AcrIF14 的 N 端结构域内的正电荷斑对于非序列特异性 dsDNA 与 Csy-AcrIF14 复合物的结合至关重要，这与 AcrIF9 的机制不同。我们的发现强调了 Acr 诱导的非特异性 DNA 结合的普遍性，并为未来研究此类 Acr 蛋白的机制提供了线索。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ee4/8464039/1bd977a24a96/gkab738fig7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ee4/8464039/c98c8bcd7d87/gkab738fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ee4/8464039/d0ed7616642f/gkab738fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ee4/8464039/98860af9f347/gkab738fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ee4/8464039/20a1c9ae918a/gkab738fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ee4/8464039/6a74081ca77b/gkab738fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ee4/8464039/16b1962b9228/gkab738fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ee4/8464039/1bd977a24a96/gkab738fig7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ee4/8464039/c98c8bcd7d87/gkab738fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ee4/8464039/d0ed7616642f/gkab738fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ee4/8464039/98860af9f347/gkab738fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ee4/8464039/20a1c9ae918a/gkab738fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ee4/8464039/6a74081ca77b/gkab738fig5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ee4/8464039/16b1962b9228/gkab738fig6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ee4/8464039/1bd977a24a96/gkab738fig7.jpg

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2
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3
Discovery of multiple anti-CRISPRs highlights anti-defense gene clustering in mobile genetic elements.
AcrIF24 作为一种抗 CRISPR 蛋白和转录抑制剂的结构基础。
Nat Chem Biol. 2022 Dec;18(12):1417-1424. doi: 10.1038/s41589-022-01137-w. Epub 2022 Sep 26.
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Disarming of type I-F CRISPR-Cas surveillance complex by anti-CRISPR proteins AcrIF6 and AcrIF9.I 型 CRISPR-Cas 监视复合物的失活由抗 CRISPR 蛋白 AcrIF6 和 AcrIF9 实现。
Sci Rep. 2022 Sep 15;12(1):15548. doi: 10.1038/s41598-022-19797-y.
发现多种抗 CRISPR 蛋白突显了移动遗传元件中抗防御基因的聚类。
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