Structural Investigation of the Anti-CRISPR Protein AcrIE7.

IF 3.2 4区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Proteins-Structure Function and Bioinformatics Pub Date : 2025-05-03 DOI:10.1002/prot.26832

Jeehee Kang, Changkon Park, Gyujin Lee, Jasung Koo, Hyejin Oh, Eun-Hee Kim, Euiyoung Bae, Jeong-Yong Suh

引用次数: 0

Abstract

The CRISPR-Cas system is an adaptive immune system in prokaryotes that provides protection against bacteriophages. As a countermeasure, bacteriophages have evolved various anti-CRISPR proteins that neutralize CRISPR-Cas immunity. Here, we report the structural and functional investigation of AcrIE7, which inhibits the type I-E CRISPR-Cas system in Pseudomonas aeruginosa. We determined both crystal and solution structures of AcrIE7, which revealed a novel helical fold. In binding assays using various biochemical methods, AcrIE7 did not tightly interact with a single Cas component in the type I-E Cascade complex or the CRISPR adaptation machinery. In contrast, AlphaFold modeling with our experimentally determined AcrIE7 structure predicted that AcrIE7 interacts with Cas3 in the type I-E CRISPR-Cas system in P. aeruginosa. Our findings are consistent with a model where AcrIE7 inhibits Cas3 and also highlight the effectiveness and limitations of AlphaFold modeling.

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抗crispr蛋白AcrIE7的结构研究

CRISPR-Cas系统是原核生物中的一种适应性免疫系统，可提供对噬菌体的保护。作为对策，噬菌体进化出各种抗crispr蛋白来中和CRISPR-Cas免疫。在这里，我们报道了抑制铜绿假单胞菌I-E型CRISPR-Cas系统的AcrIE7的结构和功能研究。我们测定了AcrIE7的晶体和溶液结构，揭示了一种新的螺旋褶皱。在使用各种生化方法的结合试验中，AcrIE7不与I-E型级联复合体或CRISPR适应机制中的单个Cas组分紧密相互作用。相比之下，使用我们实验确定的AcrIE7结构的AlphaFold模型预测，在铜绿假单胞菌的I-E型CRISPR-Cas系统中，AcrIE7与Cas3相互作用。我们的发现与AcrIE7抑制Cas3的模型一致，也突出了AlphaFold模型的有效性和局限性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Proteins-Structure Function and Bioinformatics 生物-生化与分子生物学

CiteScore

5.90

自引率

3.40%

发文量

172

审稿时长

3 months

期刊介绍： PROTEINS : Structure, Function, and Bioinformatics publishes original reports of significant experimental and analytic research in all areas of protein research: structure, function, computation, genetics, and design. The journal encourages reports that present new experimental or computational approaches for interpreting and understanding data from biophysical chemistry, structural studies of proteins and macromolecular assemblies, alterations of protein structure and function engineered through techniques of molecular biology and genetics, functional analyses under physiologic conditions, as well as the interactions of proteins with receptors, nucleic acids, or other specific ligands or substrates. Research in protein and peptide biochemistry directed toward synthesizing or characterizing molecules that simulate aspects of the activity of proteins, or that act as inhibitors of protein function, is also within the scope of PROTEINS. In addition to full-length reports, short communications (usually not more than 4 printed pages) and prediction reports are welcome. Reviews are typically by invitation; authors are encouraged to submit proposed topics for consideration.