Hyper-stimulation of Pyrococcus furiosus CRISPR DNA uptake by a self-transmissible plasmid.

Autor: Watts EA; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602, USA., Garrett SC; Department of Genetics and Genome Sciences, Institute for Systems Genomics, University of Connecticut Health Center, Farmington, CT, 06030, USA., Catchpole RJ; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602, USA., Clark LM; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602, USA., Graveley BR; Department of Genetics and Genome Sciences, Institute for Systems Genomics, University of Connecticut Health Center, Farmington, CT, 06030, USA. graveley@uchc.edu., Terns MP; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602, USA. mterns@uga.edu.; Department of Genetics, University of Georgia, Athens, GA, 30602, USA. mterns@uga.edu.; Department of Microbiology, University of Georgia, Athens, GA, 30602, USA. mterns@uga.edu.
Jazyk: angličtina
Zdroj: Extremophiles : life under extreme conditions [Extremophiles] 2022 Nov 16; Vol. 26 (3), pp. 36. Date of Electronic Publication: 2022 Nov 16.
DOI: 10.1007/s00792-022-01281-0
Abstrakt: Pyrococcus furiosus is a hyperthermophilic archaeon with three effector CRISPR complexes (types I-A, I-B, and III-B) that each employ crRNAs derived from seven CRISPR arrays. Here, we investigate the CRISPR adaptation response to a newly discovered and self-transmissible plasmid, pT33.3. Transconjugant strains of Pyrococcus furiosus exhibited dramatically elevated levels of new spacer integration at CRISPR loci relative to the strain harboring a commonly employed, laboratory-constructed plasmid. High-throughput sequence analysis demonstrated that the vast majority of the newly acquired spacers were preferentially selected from DNA surrounding a particular region of the pT33.3 plasmid and exhibited a bi-directional pattern of strand bias that is a hallmark of primed adaptation by type I systems. We observed that one of the CRISPR arrays of our Pyrococcus furiosus laboratory strain encodes a spacer that closely matches the region of the conjugative plasmid that is targeted for adaptation. The hyper-adaptation phenotype was found to strictly depend both on the presence of this single matching spacer as well as the I-B effector complex, known to mediate primed adaptation. Our results indicate that Pyrococcus furiosus naturally encountered this conjugative plasmid or a related mobile genetic element in the past and responds to reinfection with robust primed adaptation.
(© 2022. The Author(s), under exclusive licence to Springer Japan KK, part of Springer Nature.)
Databáze: MEDLINE
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