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CRISPR-Cas Controls Cryptic Prophages

Sooyeon Song, View ORCID ProfileThomas K. Wood
doi: https://doi.org/10.1101/2021.07.28.454074
Sooyeon Song
1Department of Chemical Engineering, Pennsylvania State University, University Park, Pennsylvania, 16802-4400, USA
2Departments of Animal Science, Jeonbuk National University, 587 Baekje-Daero, Deojin-Gu, Jeonju-Si, Jellabuk-Do, 54896, South Korea
3Agricultural Convergence Technology, Jeonbuk National University, 587 Baekje-Daero, Deojin-Gu, Jeonju-Si, Jellabuk-Do, 54896, South Korea
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  • For correspondence: songsy@jbnu.ac.kr tuw14@psu.edu
Thomas K. Wood
1Department of Chemical Engineering, Pennsylvania State University, University Park, Pennsylvania, 16802-4400, USA
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  • ORCID record for Thomas K. Wood
  • For correspondence: songsy@jbnu.ac.kr tuw14@psu.edu
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ABSTRACT

The bacterial archetypal adaptive immune system, CRISPR-Cas, is thought to be non-functional in the best-studied bacterium, Escherichia coli K-12. Instead, we demonstrate here that the E. coli CRISPR-Cas system is active and inhibits its nine defective (i.e., cryptic) prophages. Specifically, deactivation of CRISPR-Cas via deletion of cas2, which encodes one of the two conserved CRISPR-Cas proteins, reduces growth by 40%, increases cell death by 700%, and prevents persister cell resuscitation; hence, CRISPR-Cas serves to inhibit the remaining deleterious effects of these cryptic prophages. Consistently, seven of the 13 E. coli spacers contain matches to the cryptic prophages, and, after excision, CRISPR-Cas cleaves cryptic prophage CP4-57 and DLP-12 DNA. Moreover, we determine that the key genes in these cryptic prophages that CRISPR-Cas represses by cleaving the excised DNA include lysis protein YdfD of Qin and lysis protein RzoD of DLP-12. Therefore, we report the novel results that (i) CRISPR-Cas is active in E. coli and (ii) CRISPR-Cas is used to tame cryptic prophages; i.e., unlike with active lysogens, CRISPR-Cas and cryptic prophages may stably exist.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-ND 4.0 International license.
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Posted July 28, 2021.
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CRISPR-Cas Controls Cryptic Prophages
Sooyeon Song, Thomas K. Wood
bioRxiv 2021.07.28.454074; doi: https://doi.org/10.1101/2021.07.28.454074
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CRISPR-Cas Controls Cryptic Prophages
Sooyeon Song, Thomas K. Wood
bioRxiv 2021.07.28.454074; doi: https://doi.org/10.1101/2021.07.28.454074

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