Disabling a Type I-E CRISPR-Cas Nuclease with a Bacteriophage-Encoded Anti-CRISPR Protein

ABSTRACT CRISPR (clustered regularly interspaced short palindromic repeat)-Cas adaptive immune systems are prevalent defense mechanisms in bacteria and archaea. They provide sequence-specific detection and neutralization of foreign nucleic acids such as bacteriophages and plasmids. One mechanism by...

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Autores principales:	April Pawluk, Megha Shah, Marios Mejdani, Charles Calmettes, Trevor F. Moraes, Alan R. Davidson, Karen L. Maxwell
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	CRISPR-Cas Pseudomonas aeruginosa X-ray crystallography anti-CRISPR type I-E CRISPR-Cas Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/638a3e556a714bbc9c1150d62a69a2df
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spelling	oai:doaj.org-article:638a3e556a714bbc9c1150d62a69a2df2021-11-15T15:51:55ZDisabling a Type I-E CRISPR-Cas Nuclease with a Bacteriophage-Encoded Anti-CRISPR Protein10.1128/mBio.01751-172150-7511https://doaj.org/article/638a3e556a714bbc9c1150d62a69a2df2017-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01751-17https://doaj.org/toc/2150-7511ABSTRACT CRISPR (clustered regularly interspaced short palindromic repeat)-Cas adaptive immune systems are prevalent defense mechanisms in bacteria and archaea. They provide sequence-specific detection and neutralization of foreign nucleic acids such as bacteriophages and plasmids. One mechanism by which phages and other mobile genetic elements are able to overcome the CRISPR-Cas system is through the expression of anti-CRISPR proteins. Over 20 different families of anti-CRISPR proteins have been described, each of which inhibits a particular type of CRISPR-Cas system. In this work, we determined the structure of type I-E anti-CRISPR protein AcrE1 by X-ray crystallography. We show that AcrE1 binds to the CRISPR-associated helicase/nuclease Cas3 and that the C-terminal region of the anti-CRISPR protein is important for its inhibitory activity. We further show that AcrE1 can convert the endogenous type I-E CRISPR system into a programmable transcriptional repressor. IMPORTANCE The CRISPR-Cas immune system provides bacteria with resistance to invasion by potentially harmful viruses, plasmids, and other foreign mobile genetic elements. This study presents the first structural and mechanistic insight into a phage-encoded protein that inactivates the type I-E CRISPR-Cas system in Pseudomonas aeruginosa. The interaction of this anti-CRISPR protein with the CRISPR-associated helicase/nuclease proteins Cas3 shuts down the CRISPR-Cas system and protects phages carrying this gene from destruction. This interaction also allows the repurposing of the endogenous type I-E CRISPR system into a programmable transcriptional repressor, providing a new biotechnological tool for genetic studies of bacteria encoding this type I-E CRISPR-Cas system.April PawlukMegha ShahMarios MejdaniCharles CalmettesTrevor F. MoraesAlan R. DavidsonKaren L. MaxwellAmerican Society for MicrobiologyarticleCRISPR-CasPseudomonas aeruginosaX-ray crystallographyanti-CRISPRtype I-E CRISPR-CasMicrobiologyQR1-502ENmBio, Vol 8, Iss 6 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	CRISPR-Cas Pseudomonas aeruginosa X-ray crystallography anti-CRISPR type I-E CRISPR-Cas Microbiology QR1-502
spellingShingle	CRISPR-Cas Pseudomonas aeruginosa X-ray crystallography anti-CRISPR type I-E CRISPR-Cas Microbiology QR1-502 April Pawluk Megha Shah Marios Mejdani Charles Calmettes Trevor F. Moraes Alan R. Davidson Karen L. Maxwell Disabling a Type I-E CRISPR-Cas Nuclease with a Bacteriophage-Encoded Anti-CRISPR Protein
description	ABSTRACT CRISPR (clustered regularly interspaced short palindromic repeat)-Cas adaptive immune systems are prevalent defense mechanisms in bacteria and archaea. They provide sequence-specific detection and neutralization of foreign nucleic acids such as bacteriophages and plasmids. One mechanism by which phages and other mobile genetic elements are able to overcome the CRISPR-Cas system is through the expression of anti-CRISPR proteins. Over 20 different families of anti-CRISPR proteins have been described, each of which inhibits a particular type of CRISPR-Cas system. In this work, we determined the structure of type I-E anti-CRISPR protein AcrE1 by X-ray crystallography. We show that AcrE1 binds to the CRISPR-associated helicase/nuclease Cas3 and that the C-terminal region of the anti-CRISPR protein is important for its inhibitory activity. We further show that AcrE1 can convert the endogenous type I-E CRISPR system into a programmable transcriptional repressor. IMPORTANCE The CRISPR-Cas immune system provides bacteria with resistance to invasion by potentially harmful viruses, plasmids, and other foreign mobile genetic elements. This study presents the first structural and mechanistic insight into a phage-encoded protein that inactivates the type I-E CRISPR-Cas system in Pseudomonas aeruginosa. The interaction of this anti-CRISPR protein with the CRISPR-associated helicase/nuclease proteins Cas3 shuts down the CRISPR-Cas system and protects phages carrying this gene from destruction. This interaction also allows the repurposing of the endogenous type I-E CRISPR system into a programmable transcriptional repressor, providing a new biotechnological tool for genetic studies of bacteria encoding this type I-E CRISPR-Cas system.
format	article
author	April Pawluk Megha Shah Marios Mejdani Charles Calmettes Trevor F. Moraes Alan R. Davidson Karen L. Maxwell
author_facet	April Pawluk Megha Shah Marios Mejdani Charles Calmettes Trevor F. Moraes Alan R. Davidson Karen L. Maxwell
author_sort	April Pawluk
title	Disabling a Type I-E CRISPR-Cas Nuclease with a Bacteriophage-Encoded Anti-CRISPR Protein
title_short	Disabling a Type I-E CRISPR-Cas Nuclease with a Bacteriophage-Encoded Anti-CRISPR Protein
title_full	Disabling a Type I-E CRISPR-Cas Nuclease with a Bacteriophage-Encoded Anti-CRISPR Protein
title_fullStr	Disabling a Type I-E CRISPR-Cas Nuclease with a Bacteriophage-Encoded Anti-CRISPR Protein
title_full_unstemmed	Disabling a Type I-E CRISPR-Cas Nuclease with a Bacteriophage-Encoded Anti-CRISPR Protein
title_sort	disabling a type i-e crispr-cas nuclease with a bacteriophage-encoded anti-crispr protein
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/638a3e556a714bbc9c1150d62a69a2df
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Disabling a Type I-E CRISPR-Cas Nuclease with a Bacteriophage-Encoded Anti-CRISPR Protein

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