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Publication :
Lactococcus lactis type III-A CRISPR-Cas system cleaves bacteriophage RNA

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Date

2018-10-02

Direction de publication

Direction de recherche

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Éditeur

Tandfonline

Projets de recherche

Structures organisationnelles

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Résumé

CRISPR-Cas defends microbial cells against invading nucleic acids including viral genomes. Recent studies have shown that type III-A CRISPR-Cas systems target both RNA and DNA in a transcriptiondependent manner. We previously found a type III-A system on a conjugative plasmid in Lactococcus lactis which provided resistance against virulent phages of the Siphoviridae family. Its naturally occurring spacers are oriented to generate crRNAs complementary to target phage mRNA, suggesting transcription-dependent targeting. Here, we show that only constructs whose spacers produce crRNAs complementary to the phage mRNA confer phage resistance in L. lactis. In vivo nucleic acid cleavage assays showed that cleavage of phage dsDNA genome was not detected within phage-infected L. lactis cells. On the other hand, Northern blots indicated that the lactococcal CRISPR-Cas cleaves phage mRNA in vivo. These results cannot exclude that single-stranded phage DNA is not being targeted, but phage DNA replication has been shown to be impaired.

Description

Revue

RNA Biology, Vol. 16 (4), 461–468 (2019)

DOI

10.1080/15476286.2018.1502589

URL vers la version publiée

Mots-clés

Phages, CRISPR, Lactococcus, Resistance

Citation

Licence CC

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