Prespacers formed during primed adaptation associate with the Cas1–Cas2 adaptation complex and the Cas3 interference nuclease–helicase
Open Access
- 25 May 2021
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 118 (22)
- https://doi.org/10.1073/pnas.2021291118
Abstract
For Type I CRISPR-Cas systems, a mode of CRISPR adaptation named priming has been described. Priming allows specific and highly efficient acquisition of new spacers from DNA recognized (primed) by the Cascade-crRNA (CRISPR RNA) effector complex. Recognition of the priming protospacer by Cascade-crRNA serves as a signal for engaging the Cas3 nuclease–helicase required for both interference and primed adaptation, suggesting the existence of a primed adaptation complex (PAC) containing the Cas1–Cas2 adaptation integrase and Cas3. To detect this complex in vivo, we here performed chromatin immunoprecipitation with Cas3-specific and Cas1-specific antibodies using cells undergoing primed adaptation. We found that prespacers are bound by both Cas1 (presumably, as part of the Cas1–Cas2 integrase) and Cas3, implying direct physical association of the interference and adaptation machineries as part of PAC.This publication has 72 references indexed in Scilit:
- The Population and Evolutionary Dynamics of Phage and Bacteria with CRISPR–Mediated ImmunityPLoS Genetics, 2013
- CRISPR Immunity Relies on the Consecutive Binding and Degradation of Negatively Supercoiled Invader DNA by Cascade and Cas3Molecular Cell, 2012
- Fast gapped-read alignment with Bowtie 2Nature Methods, 2012
- Proteins and DNA elements essential for the CRISPR adaptation process in Escherichia coliNucleic Acids Research, 2012
- Interference by clustered regularly interspaced short palindromic repeat (CRISPR) RNA is governed by a seed sequenceProceedings of the National Academy of Sciences of the United States of America, 2011
- Andromeda: A Peptide Search Engine Integrated into the MaxQuant EnvironmentJournal of Proteome Research, 2011
- Direct and indirect effects of H-NS and Fis on global gene expression control in Escherichia coliNucleic Acids Research, 2010
- CRISPR Interference Limits Horizontal Gene Transfer in Staphylococci by Targeting DNAScience, 2008
- MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantificationNature Biotechnology, 2008
- Small CRISPR RNAs Guide Antiviral Defense in ProkaryotesScience, 2008