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The structure of a Type III-A CRISPR-Cas effector complex reveals conserved and idiosyncratic contacts to target RNA and crRNA among Type III-A systems

Type III CRISPR-Cas systems employ multiprotein effector complexes bound to small CRISPR RNAs (crRNAs) to detect foreign RNA transcripts and elicit a complex immune response that leads to the destruction of invading RNA and DNA. Type III systems are among the most widespread in nature, and emerging...

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Autores principales: Paraan, Mohammadreza, Nasef, Mohamed, Chou-Zheng, Lucy, Khweis, Sarah A., Schoeffler, Allyn J., Hatoum-Aslan, Asma, Stagg, Scott M., Dunkle, Jack A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10289348/
https://www.ncbi.nlm.nih.gov/pubmed/37352230
http://dx.doi.org/10.1371/journal.pone.0287461
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author Paraan, Mohammadreza
Nasef, Mohamed
Chou-Zheng, Lucy
Khweis, Sarah A.
Schoeffler, Allyn J.
Hatoum-Aslan, Asma
Stagg, Scott M.
Dunkle, Jack A.
author_facet Paraan, Mohammadreza
Nasef, Mohamed
Chou-Zheng, Lucy
Khweis, Sarah A.
Schoeffler, Allyn J.
Hatoum-Aslan, Asma
Stagg, Scott M.
Dunkle, Jack A.
author_sort Paraan, Mohammadreza
collection PubMed
description Type III CRISPR-Cas systems employ multiprotein effector complexes bound to small CRISPR RNAs (crRNAs) to detect foreign RNA transcripts and elicit a complex immune response that leads to the destruction of invading RNA and DNA. Type III systems are among the most widespread in nature, and emerging interest in harnessing these systems for biotechnology applications highlights the need for detailed structural analyses of representatives from diverse organisms. We performed cryo-EM reconstructions of the Type III-A Cas10-Csm effector complex from S. epidermidis bound to an intact, cognate target RNA and identified two oligomeric states, a 276 kDa complex and a 318 kDa complex. 3.1 Å density for the well-ordered 276 kDa complex allowed construction of atomic models for the Csm2, Csm3, Csm4 and Csm5 subunits within the complex along with the crRNA and target RNA. We also collected small-angle X-ray scattering data which was consistent with the 276 kDa Cas10-Csm architecture we identified. Detailed comparisons between the S. epidermidis Cas10-Csm structure and the well-resolved bacterial (S. thermophilus) and archaeal (T. onnurineus) Cas10-Csm structures reveal differences in how the complexes interact with target RNA and crRNA which are likely to have functional ramifications. These structural comparisons shed light on the unique features of Type III-A systems from diverse organisms and will assist in improving biotechnologies derived from Type III-A effector complexes.
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spelling pubmed-102893482023-06-24 The structure of a Type III-A CRISPR-Cas effector complex reveals conserved and idiosyncratic contacts to target RNA and crRNA among Type III-A systems Paraan, Mohammadreza Nasef, Mohamed Chou-Zheng, Lucy Khweis, Sarah A. Schoeffler, Allyn J. Hatoum-Aslan, Asma Stagg, Scott M. Dunkle, Jack A. PLoS One Research Article Type III CRISPR-Cas systems employ multiprotein effector complexes bound to small CRISPR RNAs (crRNAs) to detect foreign RNA transcripts and elicit a complex immune response that leads to the destruction of invading RNA and DNA. Type III systems are among the most widespread in nature, and emerging interest in harnessing these systems for biotechnology applications highlights the need for detailed structural analyses of representatives from diverse organisms. We performed cryo-EM reconstructions of the Type III-A Cas10-Csm effector complex from S. epidermidis bound to an intact, cognate target RNA and identified two oligomeric states, a 276 kDa complex and a 318 kDa complex. 3.1 Å density for the well-ordered 276 kDa complex allowed construction of atomic models for the Csm2, Csm3, Csm4 and Csm5 subunits within the complex along with the crRNA and target RNA. We also collected small-angle X-ray scattering data which was consistent with the 276 kDa Cas10-Csm architecture we identified. Detailed comparisons between the S. epidermidis Cas10-Csm structure and the well-resolved bacterial (S. thermophilus) and archaeal (T. onnurineus) Cas10-Csm structures reveal differences in how the complexes interact with target RNA and crRNA which are likely to have functional ramifications. These structural comparisons shed light on the unique features of Type III-A systems from diverse organisms and will assist in improving biotechnologies derived from Type III-A effector complexes. Public Library of Science 2023-06-23 /pmc/articles/PMC10289348/ /pubmed/37352230 http://dx.doi.org/10.1371/journal.pone.0287461 Text en © 2023 Paraan et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Paraan, Mohammadreza
Nasef, Mohamed
Chou-Zheng, Lucy
Khweis, Sarah A.
Schoeffler, Allyn J.
Hatoum-Aslan, Asma
Stagg, Scott M.
Dunkle, Jack A.
The structure of a Type III-A CRISPR-Cas effector complex reveals conserved and idiosyncratic contacts to target RNA and crRNA among Type III-A systems
title The structure of a Type III-A CRISPR-Cas effector complex reveals conserved and idiosyncratic contacts to target RNA and crRNA among Type III-A systems
title_full The structure of a Type III-A CRISPR-Cas effector complex reveals conserved and idiosyncratic contacts to target RNA and crRNA among Type III-A systems
title_fullStr The structure of a Type III-A CRISPR-Cas effector complex reveals conserved and idiosyncratic contacts to target RNA and crRNA among Type III-A systems
title_full_unstemmed The structure of a Type III-A CRISPR-Cas effector complex reveals conserved and idiosyncratic contacts to target RNA and crRNA among Type III-A systems
title_short The structure of a Type III-A CRISPR-Cas effector complex reveals conserved and idiosyncratic contacts to target RNA and crRNA among Type III-A systems
title_sort structure of a type iii-a crispr-cas effector complex reveals conserved and idiosyncratic contacts to target rna and crrna among type iii-a systems
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10289348/
https://www.ncbi.nlm.nih.gov/pubmed/37352230
http://dx.doi.org/10.1371/journal.pone.0287461
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