Intrinsic sequence specificity of the Cas1 integrase directs new spacer acquisition

eLife

Malcolm F White¹  Clare Rollie¹  Stefanie Schneider²  Edward L Bolt³  Anna Sophie Brinkmann³ 

[1] Biomedical Sciences Research Complex, University of St Andrews, St Andrews, United Kingdom;Faculty of Medicine, Institute of Cell Biology, University of Duisburg-Essen, Essen, Germany;School of Life Sciences, Queen's Medical Centre, University of Nottingham, Nottingham, United Kingdom;

关键词: Sulfolobus solfataricus;    CRISPR;    integrase;    adaptation;   

DOI  :  10.7554/eLife.08716

来源: DOAJ

【 摘 要 】

The adaptive prokaryotic immune system CRISPR-Cas provides RNA-mediated protection from invading genetic elements. The fundamental basis of the system is the ability to capture small pieces of foreign DNA for incorporation into the genome at the CRISPR locus, a process known as Adaptation, which is dependent on the Cas1 and Cas2 proteins. We demonstrate that Cas1 catalyses an efficient trans-esterification reaction on branched DNA substrates, which represents the reverse- or disintegration reaction. Cas1 from both Escherichia coli and Sulfolobus solfataricus display sequence specific activity, with a clear preference for the nucleotides flanking the integration site at the leader-repeat 1 boundary of the CRISPR locus. Cas2 is not required for this activity and does not influence the specificity. This suggests that the inherent sequence specificity of Cas1 is a major determinant of the adaptation process.

【 授权许可】

Unknown