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DNA targeting by the type I-G and type I-A CRISPR–Cas systems of Pyrococcus furiosus
CRISPR–Cas systems silence plasmids and viruses in prokaryotes. CRISPR–Cas effector complexes contain CRISPR RNAs (crRNAs) that include sequences captured from invaders and direct CRISPR-associated (Cas) proteins to destroy corresponding invader nucleic acids. Pyrococcus furiosus (Pfu) harbors three...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4666368/ https://www.ncbi.nlm.nih.gov/pubmed/26519471 http://dx.doi.org/10.1093/nar/gkv1140 |
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author | Elmore, Joshua Deighan, Trace Westpheling, Jan Terns, Rebecca M. Terns, Michael P. |
author_facet | Elmore, Joshua Deighan, Trace Westpheling, Jan Terns, Rebecca M. Terns, Michael P. |
author_sort | Elmore, Joshua |
collection | PubMed |
description | CRISPR–Cas systems silence plasmids and viruses in prokaryotes. CRISPR–Cas effector complexes contain CRISPR RNAs (crRNAs) that include sequences captured from invaders and direct CRISPR-associated (Cas) proteins to destroy corresponding invader nucleic acids. Pyrococcus furiosus (Pfu) harbors three CRISPR–Cas immune systems: a Cst (Type I-G) system with an associated Cmr (Type III-B) module at one locus, and a partial Csa (Type I-A) module (lacking known invader sequence acquisition and crRNA processing genes) at another locus. The Pfu Cmr complex cleaves complementary target RNAs, and Csa systems have been shown to target DNA, while the mechanism by which Cst complexes silence invaders is unknown. In this study, we investigated the function of the Cst as well as Csa system in Pfu strains harboring a single CRISPR–Cas system. Plasmid transformation assays revealed that the Cst and Csa systems both function by DNA silencing and utilize similar flanking sequence information (PAMs) to identify invader DNA. Silencing by each system specifically requires its associated Cas3 nuclease. crRNAs from the 7 shared CRISPR loci in Pfu are processed for use by all 3 effector complexes, and Northern analysis revealed that individual effector complexes dictate the profile of mature crRNA species that is generated. |
format | Online Article Text |
id | pubmed-4666368 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-46663682015-12-02 DNA targeting by the type I-G and type I-A CRISPR–Cas systems of Pyrococcus furiosus Elmore, Joshua Deighan, Trace Westpheling, Jan Terns, Rebecca M. Terns, Michael P. Nucleic Acids Res Molecular Biology CRISPR–Cas systems silence plasmids and viruses in prokaryotes. CRISPR–Cas effector complexes contain CRISPR RNAs (crRNAs) that include sequences captured from invaders and direct CRISPR-associated (Cas) proteins to destroy corresponding invader nucleic acids. Pyrococcus furiosus (Pfu) harbors three CRISPR–Cas immune systems: a Cst (Type I-G) system with an associated Cmr (Type III-B) module at one locus, and a partial Csa (Type I-A) module (lacking known invader sequence acquisition and crRNA processing genes) at another locus. The Pfu Cmr complex cleaves complementary target RNAs, and Csa systems have been shown to target DNA, while the mechanism by which Cst complexes silence invaders is unknown. In this study, we investigated the function of the Cst as well as Csa system in Pfu strains harboring a single CRISPR–Cas system. Plasmid transformation assays revealed that the Cst and Csa systems both function by DNA silencing and utilize similar flanking sequence information (PAMs) to identify invader DNA. Silencing by each system specifically requires its associated Cas3 nuclease. crRNAs from the 7 shared CRISPR loci in Pfu are processed for use by all 3 effector complexes, and Northern analysis revealed that individual effector complexes dictate the profile of mature crRNA species that is generated. Oxford University Press 2015-12-02 2015-10-30 /pmc/articles/PMC4666368/ /pubmed/26519471 http://dx.doi.org/10.1093/nar/gkv1140 Text en © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Molecular Biology Elmore, Joshua Deighan, Trace Westpheling, Jan Terns, Rebecca M. Terns, Michael P. DNA targeting by the type I-G and type I-A CRISPR–Cas systems of Pyrococcus furiosus |
title | DNA targeting by the type I-G and type I-A CRISPR–Cas systems of Pyrococcus furiosus |
title_full | DNA targeting by the type I-G and type I-A CRISPR–Cas systems of Pyrococcus furiosus |
title_fullStr | DNA targeting by the type I-G and type I-A CRISPR–Cas systems of Pyrococcus furiosus |
title_full_unstemmed | DNA targeting by the type I-G and type I-A CRISPR–Cas systems of Pyrococcus furiosus |
title_short | DNA targeting by the type I-G and type I-A CRISPR–Cas systems of Pyrococcus furiosus |
title_sort | dna targeting by the type i-g and type i-a crispr–cas systems of pyrococcus furiosus |
topic | Molecular Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4666368/ https://www.ncbi.nlm.nih.gov/pubmed/26519471 http://dx.doi.org/10.1093/nar/gkv1140 |
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