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Spatiotemporal Control of Type III-A CRISPR-Cas Immunity: Coupling DNA Degradation with the Target RNA Recognition

Kazlauskiene, Migle, Tamulaitis, Gintautas, Kostiuk, Georgij, Venclovas, Česlovas, Siksnys, Virginijus·2016·Molecular Cell
DOI 10.1016/j.molcel.2016.03.024
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Abstract

Streptococcus thermophilus (St) type III-A CRISPR-Cas system restricts MS2 RNA phage and cuts RNA in vitro. However, the CRISPR array spacers match DNA phages, raising the question: does the St CRISPR-Cas system provide immunity by erasing phage mRNA or/and by eliminating invading DNA? We show that it does both. We find that (1) base-pairing between crRNA and target RNA activates single-stranded DNA (ssDNA) degradation by StCsm; (2) ssDNase activity is confined to the HD-domain of Cas10; (3) target RNA cleavage by the Csm3 RNase suppresses Cas10 DNase activity, ensuring temporal control of DNA degradation; and (4) base-pairing between crRNA 5'-handle and target RNA 3'-flanking sequence inhibits Cas10 ssDNase to prevent self-targeting. We propose that upon phage infection, crRNA-guided StCsm binding to the emerging transcript recruits Cas10 DNase to the actively transcribed phage DNA, resulting in degradation of both the transcript and phage DNA, but not the host DNA.

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  • Type III-A and type III-B share the signature protein Cas10 and are unique in relation to other interference mechanisms because they target both RNA and DNA substrates.

    “Type III-A and type III-B share in common the signature protein of type III systems, Cas10, and are unique in relation to other interference mechanisms because they target both RNA and DNA substrates [48,49].”

    From CRISPR-Cas: Converting A Bacterial Defence Mechanism into A State-of-the-Art Genetic Manipulation Tool (2019) · cites this paper as [49]

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Metadata

Authors
Kazlauskiene, M., Tamulaitis, G., Kostiuk, G., Venclovas, Č., & Siksnys, V.
Venue
Molecular Cell
Publisher
Elsevier BV
Published
2016-04
Volume
62
Issue
2
Pages
295-306
ISSN
1097-2765
DOI
10.1016/j.molcel.2016.03.024
Cited by
265
References
1
Language
en
Licence
Funders
Lietuvos Mokslo Taryba (MIP-40/2013)

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