The novel anti-CRISPR AcrIIA22 relieves DNA torsion in target plasmids and impairs SpyCas9 activity.

To overcome CRISPR-Cas defense systems, many phages and mobile genetic elements (MGEs) encode CRISPR-Cas inhibitors called anti-CRISPRs (Acrs). Nearly all characterized Acrs directly bind Cas proteins to inactivate CRISPR immunity. Here, using functional metagenomic selection, we describe AcrIIA22,...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Kevin J Forsberg, Danica T Schmidtke, Rachel Werther, Ruben V Uribe, Deanna Hausman, Morten O A Sommer, Barry L Stoddard, Brett K Kaiser, Harmit S Malik
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/f4d435c0762349c6924b3c0f2c435c31
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:f4d435c0762349c6924b3c0f2c435c31
record_format	dspace
spelling	oai:doaj.org-article:f4d435c0762349c6924b3c0f2c435c312021-12-02T19:54:27ZThe novel anti-CRISPR AcrIIA22 relieves DNA torsion in target plasmids and impairs SpyCas9 activity.1544-91731545-788510.1371/journal.pbio.3001428https://doaj.org/article/f4d435c0762349c6924b3c0f2c435c312021-10-01T00:00:00Zhttps://doi.org/10.1371/journal.pbio.3001428https://doaj.org/toc/1544-9173https://doaj.org/toc/1545-7885To overcome CRISPR-Cas defense systems, many phages and mobile genetic elements (MGEs) encode CRISPR-Cas inhibitors called anti-CRISPRs (Acrs). Nearly all characterized Acrs directly bind Cas proteins to inactivate CRISPR immunity. Here, using functional metagenomic selection, we describe AcrIIA22, an unconventional Acr found in hypervariable genomic regions of clostridial bacteria and their prophages from human gut microbiomes. AcrIIA22 does not bind strongly to SpyCas9 but nonetheless potently inhibits its activity against plasmids. To gain insight into its mechanism, we obtained an X-ray crystal structure of AcrIIA22, which revealed homology to PC4-like nucleic acid-binding proteins. Based on mutational analyses and functional assays, we deduced that acrIIA22 encodes a DNA nickase that relieves torsional stress in supercoiled plasmids. This may render them less susceptible to SpyCas9, which uses free energy from negative supercoils to form stable R-loops. Modifying DNA topology may provide an additional route to CRISPR-Cas resistance in phages and MGEs.Kevin J ForsbergDanica T SchmidtkeRachel WertherRuben V UribeDeanna HausmanMorten O A SommerBarry L StoddardBrett K KaiserHarmit S MalikPublic Library of Science (PLoS)articleBiology (General)QH301-705.5ENPLoS Biology, Vol 19, Iss 10, p e3001428 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Biology (General) QH301-705.5
spellingShingle	Biology (General) QH301-705.5 Kevin J Forsberg Danica T Schmidtke Rachel Werther Ruben V Uribe Deanna Hausman Morten O A Sommer Barry L Stoddard Brett K Kaiser Harmit S Malik The novel anti-CRISPR AcrIIA22 relieves DNA torsion in target plasmids and impairs SpyCas9 activity.
description	To overcome CRISPR-Cas defense systems, many phages and mobile genetic elements (MGEs) encode CRISPR-Cas inhibitors called anti-CRISPRs (Acrs). Nearly all characterized Acrs directly bind Cas proteins to inactivate CRISPR immunity. Here, using functional metagenomic selection, we describe AcrIIA22, an unconventional Acr found in hypervariable genomic regions of clostridial bacteria and their prophages from human gut microbiomes. AcrIIA22 does not bind strongly to SpyCas9 but nonetheless potently inhibits its activity against plasmids. To gain insight into its mechanism, we obtained an X-ray crystal structure of AcrIIA22, which revealed homology to PC4-like nucleic acid-binding proteins. Based on mutational analyses and functional assays, we deduced that acrIIA22 encodes a DNA nickase that relieves torsional stress in supercoiled plasmids. This may render them less susceptible to SpyCas9, which uses free energy from negative supercoils to form stable R-loops. Modifying DNA topology may provide an additional route to CRISPR-Cas resistance in phages and MGEs.
format	article
author	Kevin J Forsberg Danica T Schmidtke Rachel Werther Ruben V Uribe Deanna Hausman Morten O A Sommer Barry L Stoddard Brett K Kaiser Harmit S Malik
author_facet	Kevin J Forsberg Danica T Schmidtke Rachel Werther Ruben V Uribe Deanna Hausman Morten O A Sommer Barry L Stoddard Brett K Kaiser Harmit S Malik
author_sort	Kevin J Forsberg
title	The novel anti-CRISPR AcrIIA22 relieves DNA torsion in target plasmids and impairs SpyCas9 activity.
title_short	The novel anti-CRISPR AcrIIA22 relieves DNA torsion in target plasmids and impairs SpyCas9 activity.
title_full	The novel anti-CRISPR AcrIIA22 relieves DNA torsion in target plasmids and impairs SpyCas9 activity.
title_fullStr	The novel anti-CRISPR AcrIIA22 relieves DNA torsion in target plasmids and impairs SpyCas9 activity.
title_full_unstemmed	The novel anti-CRISPR AcrIIA22 relieves DNA torsion in target plasmids and impairs SpyCas9 activity.
title_sort	novel anti-crispr acriia22 relieves dna torsion in target plasmids and impairs spycas9 activity.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/f4d435c0762349c6924b3c0f2c435c31
work_keys_str_mv	AT kevinjforsberg thenovelanticrispracriia22relievesdnatorsionintargetplasmidsandimpairsspycas9activity AT danicatschmidtke thenovelanticrispracriia22relievesdnatorsionintargetplasmidsandimpairsspycas9activity AT rachelwerther thenovelanticrispracriia22relievesdnatorsionintargetplasmidsandimpairsspycas9activity AT rubenvuribe thenovelanticrispracriia22relievesdnatorsionintargetplasmidsandimpairsspycas9activity AT deannahausman thenovelanticrispracriia22relievesdnatorsionintargetplasmidsandimpairsspycas9activity AT mortenoasommer thenovelanticrispracriia22relievesdnatorsionintargetplasmidsandimpairsspycas9activity AT barrylstoddard thenovelanticrispracriia22relievesdnatorsionintargetplasmidsandimpairsspycas9activity AT brettkkaiser thenovelanticrispracriia22relievesdnatorsionintargetplasmidsandimpairsspycas9activity AT harmitsmalik thenovelanticrispracriia22relievesdnatorsionintargetplasmidsandimpairsspycas9activity AT kevinjforsberg novelanticrispracriia22relievesdnatorsionintargetplasmidsandimpairsspycas9activity AT danicatschmidtke novelanticrispracriia22relievesdnatorsionintargetplasmidsandimpairsspycas9activity AT rachelwerther novelanticrispracriia22relievesdnatorsionintargetplasmidsandimpairsspycas9activity AT rubenvuribe novelanticrispracriia22relievesdnatorsionintargetplasmidsandimpairsspycas9activity AT deannahausman novelanticrispracriia22relievesdnatorsionintargetplasmidsandimpairsspycas9activity AT mortenoasommer novelanticrispracriia22relievesdnatorsionintargetplasmidsandimpairsspycas9activity AT barrylstoddard novelanticrispracriia22relievesdnatorsionintargetplasmidsandimpairsspycas9activity AT brettkkaiser novelanticrispracriia22relievesdnatorsionintargetplasmidsandimpairsspycas9activity AT harmitsmalik novelanticrispracriia22relievesdnatorsionintargetplasmidsandimpairsspycas9activity
_version_	1718375938944663552

The novel anti-CRISPR AcrIIA22 relieves DNA torsion in target plasmids and impairs SpyCas9 activity.

Ejemplares similares