Tamulaitis, G., Kazlauskiene, M., Manakova, E. et al. (5 more authors) (2014) Programmable RNA Shredding by the Type III-A CRISPR-Cas System of Streptococcus thermophilus. Molecular Cell , 56 (4). pp. 506-517. ISSN 1097-2765
Abstract
Immunity against viruses and plasmids provided by CRISPR-Cas systems relies on a ribonucleoprotein effector complex that triggers the degradation of invasive nucleic acids (NA). Effector complexes of type I (Cascade) and II (Cas9-dual RNA) target foreign DNA. Intriguingly, the genetic evidence suggests that the type III-A Csm complex targets DNA, whereas biochemical data show that the type III-B Cmr complex cleaves RNA. Here we aimed to investigate NA specificity and mechanism of CRISPR interference for the Streptococcus thermophilus Csm (III-A) complex (StCsm). When expressed in Escherichia coli, two complexes of different stoichiometry copurified with 40 and 72 nt crRNA species, respectively. Both complexes targeted RNA and generated multiple cuts at 6 nt intervals. The Csm3 protein, present in multiple copies in both Csm complexes, acts as endoribonuclease. In the heterologous E. coli host, StCsm restricts MS2 RNA phage in a Csm3 nuclease-dependent manner. Thus, our results demonstrate that the type III-A StCsm complex guided by crRNA targets RNA and not DNA.
Highlights • Streptococcus thermophilus type III-A Csm (StCsm) complex targets RNA
•Multiple cuts are introduced in the target RNA at 6 nt intervals
•Csm3 protein subunits are responsible for endoribonuclease activity of the complex
•StCsm complex offers a programmable tool for RNA degradation
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2014 Elsevier. This is an author produced version of a paper subsequently published in Molecular Cell. Uploaded in accordance with the publisher's self-archiving policy. Article available under the terms of the CC-BY-NC-ND licence (https://creativecommons.org/licenses/by-nc-nd/4.0/). |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Engineering (Sheffield) > Department of Chemical and Biological Engineering (Sheffield) |
Funding Information: | Funder Grant number ENGINEERING AND PHYSICAL SCIENCE RESEARCH COUNCIL (EPSRC) EP/D033713/1 |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 08 Feb 2018 16:32 |
Last Modified: | 10 Feb 2018 05:57 |
Published Version: | https://doi.org/10.1016/j.molcel.2014.09.027 |
Status: | Published |
Publisher: | Elsevier |
Refereed: | Yes |
Identification Number: | 10.1016/j.molcel.2014.09.027 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:127155 |