Interplay of recombination, plasmid stability, and CRISPR-Cas immunity in Escherichia coli (CROSBI ID 659441)
Prilog sa skupa u zborniku | sažetak izlaganja sa skupa | međunarodna recenzija
Podaci o odgovornosti
Radovčić, Marin ; Bolt, Edward L., Ivančić-Baće, Ivana
engleski
Interplay of recombination, plasmid stability, and CRISPR-Cas immunity in Escherichia coli
CRISPR-Cas system is a prokaryotic adaptive immune system against foreign genetic elements. Immunity is acquired through insertion of small fragments of invader DNA into a CRISPR array. This process of fragment (spacer) insertion is called adaptation, and can be mediated solely by the protein complex Cas1-Cas2 (“naïve CRISPR adaptation”). In E. coli, naïve adaptation is helped by RecBCD enzyme to generate single-stranded DNA intermediates. We report here that cells lacking recD gene and transformed with Cas1-Cas2 expressing plasmid could not acquire new spacers, as expected, but that this corresponded with moderate loss of pCas1-Cas2. The adaptation could be re-established by deletion of recA, but the plasmid loss remained and the cell’s viability became reduced. Since RecA loading is constitutive in recD mutant, the presence of bound RecA onto 3’ ssDNA likely prevents reannealing of ssDNA fragments while at the same time it promotes recombination that generates unstable plasmid multimers.
Cas1-Cas2, RecBCD, RecA, E. coli
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Podaci o prilogu
3-3.
2017.
objavljeno
Podaci o matičnoj publikaciji
Cell Symposia CRISPR: From biology to technology and novel therapeutics
Pham, John ; Saxe, Jon ; Joung, Keith ; May, Andy
online: Cell Press, Elsevier
Podaci o skupu
Cell Symposia CRISPR: From biology to technology and novel therapeutics
poster
22.10.2017-24.10.2017
Sitges, Španjolska