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JKMRS Volume 25, No 3, pp 39, Backbone assignment and structur...
2021년 09월 20일 / 조회수: 36

Backbone assignment and structural analysis of anti-CRISPR AcrIF7 from Pseudomonas aeruginosa prophages

 

 

Iktae Kim1,2 and Jeong-Yong Suh1,*

 

 

1Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 08826, Republic of Korea

2Department of Biochemistry and Biophysics, Texas A&M University, College Station, TX 77843, United States

 

 

 

 

Received September 12, 2021; Revised September 18, 2021; Accepted September 18, 2021

 


Abstract The CRISPR-Cas system provides adaptive immunity for bacteria and archaea against invading phages and foreign plasmids. In the Class 1 CRISPR-Cas system, multi-subunit Cas proteins assemble with crRNA to bind to DNA targets. To disarm the bacterial defense system, bacteriophages evolved anti-CRISPR (Acr) proteins that actively inhibit the host CRISPR-Cas function. Here we report the backbone resonance assignments of AcrIF7 protein that inhibits the type I-F CRISPR-Cas system of Pseudomonas aeruginosa using triple-resonance nuclear magnetic resonance spectroscopy. We employed various computational methods to predict the structure and binding interface of AcrIF7, and assessed the model with experimental data. AcrIF7 binds to Cas8f protein via flexible loop regions to inhibit target DNA binding, suggesting that conformational heterogeneity is important for the Cas–Acr interaction.

 

Keywords AcrIF7, anti-CRISPR, chemical shifts, CRISPR-Cas

 

* Address correspondence to: Jeong-Yong Suh, Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 08826, Republic of Korea. Tel: 82-2-880-4879; E-mail: jysuh@snu.ac.kr

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