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Cas1–Cas2 complex formation mediates spacer acquisition during CRISPR–Cas adaptive immunity
- Source :
- Nature structural & molecular biology, Nuñez, James K.; Kranzusch, Philip J.; Noeske, Jonas; Wright, Addison V.; Davies, Christopher W.; & Doudna, Jennifer A.(2014). Cas1–Cas2 complex formation mediates spacer acquisition during CRISPR–Cas adaptive immunity. Nature structural & molecular biology, 21(6), 528-534. UC Office of the President: Multicampus Research Programs and Initiatives (MRPI); a funding opportunity through UC Research Initiatives (UCRI). Retrieved from: http://www.escholarship.org/uc/item/5fc8631f
- Publication Year :
- 2014
-
Abstract
- The initial stage of CRISPR-Cas immunity involves the integration of foreign DNA spacer segments into the host genomic CRISPR locus. The nucleases Cas1 and Cas2 are the only proteins conserved among all CRISPR-Cas systems, yet the molecular functions of these proteins during immunity are unknown. Here we show that Cas1 and Cas2 from Escherichia coli form a stable complex that is essential for spacer acquisition and determine the 2.3-Å-resolution crystal structure of the Cas1-Cas2 complex. Mutations that perturb Cas1-Cas2 complex formation disrupt CRISPR DNA recognition and spacer acquisition in vivo. Active site mutants of Cas2, unlike those of Cas1, can still acquire new spacers, thus indicating a nonenzymatic role of Cas2 during immunity. These results reveal the universal roles of Cas1 and Cas2 and suggest a mechanism by which Cas1-Cas2 complexes specify sites of CRISPR spacer integration.
- Subjects :
- Models, Molecular
Mutant
CRISPR-Associated Proteins
Locus (genetics)
Computational biology
Biology
Adaptive Immunity
medicine.disease_cause
Crystallography, X-Ray
Article
chemistry.chemical_compound
Protein structure
Structural Biology
Immunity
Medicine and Health Sciences
medicine
Escherichia coli
CRISPR
Clustered Regularly Interspaced Short Palindromic Repeats
Molecular Biology
Genetics
Endodeoxyribonucleases
Escherichia coli Proteins
Acquired immune system
Endonucleases
Protein Structure, Tertiary
chemistry
CRISPR-Cas Systems
DNA
Subjects
Details
- Language :
- English
- ISSN :
- 15459985 and 15459993
- Volume :
- 21
- Issue :
- 6
- Database :
- OpenAIRE
- Journal :
- Nature structural & molecular biology
- Accession number :
- edsair.doi.dedup.....14f737e704cb88daa24543e21167edae