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Bipartite recognition of target RNAs activates DNA cleavage by the Type III-B CRISPR–Cas system

CRISPR–Cas systems eliminate nucleic acid invaders in bacteria and archaea. The effector complex of the Type III-B Cmr system cleaves invader RNAs recognized by the CRISPR RNA (crRNA ) of the complex. Here we show that invader RNAs also activate the Cmr complex to cleave DNA. As has been observed fo...

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Autores principales: Elmore, Joshua R., Sheppard, Nolan F., Ramia, Nancy, Deighan, Trace, Li, Hong, Terns, Rebecca M., Terns, Michael P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4762429/
https://www.ncbi.nlm.nih.gov/pubmed/26848045
http://dx.doi.org/10.1101/gad.272153.115
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author Elmore, Joshua R.
Sheppard, Nolan F.
Ramia, Nancy
Deighan, Trace
Li, Hong
Terns, Rebecca M.
Terns, Michael P.
author_facet Elmore, Joshua R.
Sheppard, Nolan F.
Ramia, Nancy
Deighan, Trace
Li, Hong
Terns, Rebecca M.
Terns, Michael P.
author_sort Elmore, Joshua R.
collection PubMed
description CRISPR–Cas systems eliminate nucleic acid invaders in bacteria and archaea. The effector complex of the Type III-B Cmr system cleaves invader RNAs recognized by the CRISPR RNA (crRNA ) of the complex. Here we show that invader RNAs also activate the Cmr complex to cleave DNA. As has been observed for other Type III systems, Cmr eliminates plasmid invaders in Pyrococcus furiosus by a mechanism that depends on transcription of the crRNA target sequence within the plasmid. Notably, we found that the target RNA per se induces DNA cleavage by the Cmr complex in vitro. DNA cleavage activity does not depend on cleavage of the target RNA but notably does require the presence of a short sequence adjacent to the target sequence within the activating target RNA (rPAM [RNA protospacer-adjacent motif]). The activated complex does not require a target sequence (or a PAM) in the DNA substrate. Plasmid elimination by the P. furiosus Cmr system also does not require the Csx1 (CRISPR-associated Rossman fold [CARF] superfamily) protein. Plasmid silencing depends on the HD nuclease and Palm domains of the Cmr2 (Cas10 superfamily) protein. The results establish the Cmr complex as a novel DNA nuclease activated by invader RNAs containing a crRNA target sequence and a rPAM.
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spelling pubmed-47624292016-08-15 Bipartite recognition of target RNAs activates DNA cleavage by the Type III-B CRISPR–Cas system Elmore, Joshua R. Sheppard, Nolan F. Ramia, Nancy Deighan, Trace Li, Hong Terns, Rebecca M. Terns, Michael P. Genes Dev Research Paper CRISPR–Cas systems eliminate nucleic acid invaders in bacteria and archaea. The effector complex of the Type III-B Cmr system cleaves invader RNAs recognized by the CRISPR RNA (crRNA ) of the complex. Here we show that invader RNAs also activate the Cmr complex to cleave DNA. As has been observed for other Type III systems, Cmr eliminates plasmid invaders in Pyrococcus furiosus by a mechanism that depends on transcription of the crRNA target sequence within the plasmid. Notably, we found that the target RNA per se induces DNA cleavage by the Cmr complex in vitro. DNA cleavage activity does not depend on cleavage of the target RNA but notably does require the presence of a short sequence adjacent to the target sequence within the activating target RNA (rPAM [RNA protospacer-adjacent motif]). The activated complex does not require a target sequence (or a PAM) in the DNA substrate. Plasmid elimination by the P. furiosus Cmr system also does not require the Csx1 (CRISPR-associated Rossman fold [CARF] superfamily) protein. Plasmid silencing depends on the HD nuclease and Palm domains of the Cmr2 (Cas10 superfamily) protein. The results establish the Cmr complex as a novel DNA nuclease activated by invader RNAs containing a crRNA target sequence and a rPAM. Cold Spring Harbor Laboratory Press 2016-02-15 /pmc/articles/PMC4762429/ /pubmed/26848045 http://dx.doi.org/10.1101/gad.272153.115 Text en © 2016 Elmore et al.; Published by Cold Spring Harbor Laboratory Press http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
spellingShingle Research Paper
Elmore, Joshua R.
Sheppard, Nolan F.
Ramia, Nancy
Deighan, Trace
Li, Hong
Terns, Rebecca M.
Terns, Michael P.
Bipartite recognition of target RNAs activates DNA cleavage by the Type III-B CRISPR–Cas system
title Bipartite recognition of target RNAs activates DNA cleavage by the Type III-B CRISPR–Cas system
title_full Bipartite recognition of target RNAs activates DNA cleavage by the Type III-B CRISPR–Cas system
title_fullStr Bipartite recognition of target RNAs activates DNA cleavage by the Type III-B CRISPR–Cas system
title_full_unstemmed Bipartite recognition of target RNAs activates DNA cleavage by the Type III-B CRISPR–Cas system
title_short Bipartite recognition of target RNAs activates DNA cleavage by the Type III-B CRISPR–Cas system
title_sort bipartite recognition of target rnas activates dna cleavage by the type iii-b crispr–cas system
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4762429/
https://www.ncbi.nlm.nih.gov/pubmed/26848045
http://dx.doi.org/10.1101/gad.272153.115
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