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The Effect of DNA Topology on Observed Rates of R-Loop Formation and DNA Strand Cleavage by CRISPR Cas12a
Open AccessArticle

Genome Maintenance Proteins Modulate Autoimmunity Mediated Primed Adaptation by the Escherichia coli Type I-E CRISPR-Cas System

1
Center of Life Sciences, Skolkovo Institute of Science and Technology, Moscow 143028, Russia
2
Waksman Institute, Rutgers, the State University of New Jersey, Piscataway, NJ 08854, USA
3
Institute of Molecular Genetics, Russian Academy of Sciences, Moscow 123182, Russia
4
Peter the Great St Petersburg State Polytechnic University, St Petersburg 195251, Russia
5
Department of Biology, Faculty of Science, University of Zagreb, Horvatovac 102a, 10000 Zagreb, Croatia
6
Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Institute of Gene Biology, Russian Academy of Sciences, 34/5 Vavilov St., Moscow 119334, Russia
*
Author to whom correspondence should be addressed.
Deceased.
Genes 2019, 10(11), 872; https://doi.org/10.3390/genes10110872
Received: 11 October 2019 / Revised: 22 October 2019 / Accepted: 28 October 2019 / Published: 31 October 2019
(This article belongs to the Special Issue CRISPR-Cas: Interactions with Genome and Physiological Maintenance)
Bacteria and archaea use CRISPR-Cas adaptive immunity systems to interfere with viruses, plasmids, and other mobile genetic elements. During the process of adaptation, CRISPR-Cas systems acquire immunity by incorporating short fragments of invaders’ genomes into CRISPR arrays. The acquisition of fragments of host genomes leads to autoimmunity and may drive chromosomal rearrangements, negative cell selection, and influence bacterial evolution. In this study, we investigated the role of proteins involved in genome stability maintenance in spacer acquisition by the Escherichia coli type I-E CRISPR-Cas system targeting its own genome. We show here, that the deletion of recJ decreases adaptation efficiency and affects accuracy of spacers incorporation into CRISPR array. Primed adaptation efficiency is also dramatically inhibited in double mutants lacking recB and sbcD but not in single mutants suggesting independent involvement and redundancy of RecBCD and SbcCD pathways in spacer acquisition. While the presence of at least one of two complexes is crucial for efficient primed adaptation, RecBCD and SbcCD affect the pattern of acquired spacers. Overall, our data suggest distinct roles of the RecBCD and SbcCD complexes and of RecJ in spacer precursor selection and insertion into CRISPR array and highlight the functional interplay between CRISPR-Cas systems and host genome maintenance mechanisms. View Full-Text
Keywords: CRISPR-Cas; Escherichia coli; primed adaptation; DNA repair CRISPR-Cas; Escherichia coli; primed adaptation; DNA repair
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Kurilovich, E.; Shiriaeva, A.; Metlitskaya, A.; Morozova, N.; Ivancic-Bace, I.; Severinov, K.; Savitskaya, E. Genome Maintenance Proteins Modulate Autoimmunity Mediated Primed Adaptation by the Escherichia coli Type I-E CRISPR-Cas System. Genes 2019, 10, 872.

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