1/20/2023 0 Comments Integration the stranded lightWe propose that disintegration-promoted integration is functionally important in the adaptive phase of CRISPR-mediated bacterial immunity, and perhaps in other analogous transposition reactions. One-ended integration intermediates may mature into complete spacer insertions via DNA repair pathways that are also involved in transposon mobility. The second insertion event is most often accompanied by the disintegration of the first, mediated either by the 3′-hydroxyl exposed during integration or by water. In linear target sites, one-ended insertions far outnumber complete protospacer insertions. In supercoiled target sites, the predominant outcome is the disintegration of one-ended insertions that fail to complete the second integration event. Here, we challenge this view with a biochemical investigation of the mechanism of protospacer insertion, which is mechanistically analogous to DNA transposition, by the Streptococcus pyogenes Cas1-Cas2 complex. ‘Disintegration’-the reversal of transposon DNA integration at a target site-is regarded as an abortive off-pathway reaction.
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