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Cas1–Cas2 complex formation mediates spacer acquisition during CRISPR–Cas adaptive immunity
by
Nuñez, James K
, Doudna, Jennifer A
, Kranzusch, Philip J
, Wright, Addison V
, Noeske, Jonas
, Davies, Christopher W
in
45/29
/ 45/70
/ 45/77
/ 631/45/535/1266
/ 82/1
/ 82/16
/ 82/80
/ 82/83
/ Adaptive Immunity
/ Biochemistry
/ Biological Microscopy
/ Clustered Regularly Interspaced Short Palindromic Repeats - physiology
/ CRISPR-Associated Proteins - chemistry
/ CRISPR-Associated Proteins - metabolism
/ CRISPR-Associated Proteins - physiology
/ CRISPR-Cas Systems
/ Crystal structure
/ Crystallography, X-Ray
/ Deoxyribonucleic acid
/ DNA
/ E coli
/ Endodeoxyribonucleases - chemistry
/ Endodeoxyribonucleases - metabolism
/ Endodeoxyribonucleases - physiology
/ Endonucleases - chemistry
/ Endonucleases - metabolism
/ Endonucleases - physiology
/ Escherichia coli - immunology
/ Escherichia coli - metabolism
/ Escherichia coli Proteins - chemistry
/ Escherichia coli Proteins - metabolism
/ Escherichia coli Proteins - physiology
/ Genetic aspects
/ Genomics
/ Immune response
/ Immune system
/ Intergenic DNA
/ Life Sciences
/ Membrane Biology
/ Models, Molecular
/ Mutation
/ Nucleases
/ Properties
/ Protein Structure
/ Protein Structure, Tertiary
/ Proteins
/ Structure
2014
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Cas1–Cas2 complex formation mediates spacer acquisition during CRISPR–Cas adaptive immunity
by
Nuñez, James K
, Doudna, Jennifer A
, Kranzusch, Philip J
, Wright, Addison V
, Noeske, Jonas
, Davies, Christopher W
in
45/29
/ 45/70
/ 45/77
/ 631/45/535/1266
/ 82/1
/ 82/16
/ 82/80
/ 82/83
/ Adaptive Immunity
/ Biochemistry
/ Biological Microscopy
/ Clustered Regularly Interspaced Short Palindromic Repeats - physiology
/ CRISPR-Associated Proteins - chemistry
/ CRISPR-Associated Proteins - metabolism
/ CRISPR-Associated Proteins - physiology
/ CRISPR-Cas Systems
/ Crystal structure
/ Crystallography, X-Ray
/ Deoxyribonucleic acid
/ DNA
/ E coli
/ Endodeoxyribonucleases - chemistry
/ Endodeoxyribonucleases - metabolism
/ Endodeoxyribonucleases - physiology
/ Endonucleases - chemistry
/ Endonucleases - metabolism
/ Endonucleases - physiology
/ Escherichia coli - immunology
/ Escherichia coli - metabolism
/ Escherichia coli Proteins - chemistry
/ Escherichia coli Proteins - metabolism
/ Escherichia coli Proteins - physiology
/ Genetic aspects
/ Genomics
/ Immune response
/ Immune system
/ Intergenic DNA
/ Life Sciences
/ Membrane Biology
/ Models, Molecular
/ Mutation
/ Nucleases
/ Properties
/ Protein Structure
/ Protein Structure, Tertiary
/ Proteins
/ Structure
2014
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Cas1–Cas2 complex formation mediates spacer acquisition during CRISPR–Cas adaptive immunity
by
Nuñez, James K
, Doudna, Jennifer A
, Kranzusch, Philip J
, Wright, Addison V
, Noeske, Jonas
, Davies, Christopher W
in
45/29
/ 45/70
/ 45/77
/ 631/45/535/1266
/ 82/1
/ 82/16
/ 82/80
/ 82/83
/ Adaptive Immunity
/ Biochemistry
/ Biological Microscopy
/ Clustered Regularly Interspaced Short Palindromic Repeats - physiology
/ CRISPR-Associated Proteins - chemistry
/ CRISPR-Associated Proteins - metabolism
/ CRISPR-Associated Proteins - physiology
/ CRISPR-Cas Systems
/ Crystal structure
/ Crystallography, X-Ray
/ Deoxyribonucleic acid
/ DNA
/ E coli
/ Endodeoxyribonucleases - chemistry
/ Endodeoxyribonucleases - metabolism
/ Endodeoxyribonucleases - physiology
/ Endonucleases - chemistry
/ Endonucleases - metabolism
/ Endonucleases - physiology
/ Escherichia coli - immunology
/ Escherichia coli - metabolism
/ Escherichia coli Proteins - chemistry
/ Escherichia coli Proteins - metabolism
/ Escherichia coli Proteins - physiology
/ Genetic aspects
/ Genomics
/ Immune response
/ Immune system
/ Intergenic DNA
/ Life Sciences
/ Membrane Biology
/ Models, Molecular
/ Mutation
/ Nucleases
/ Properties
/ Protein Structure
/ Protein Structure, Tertiary
/ Proteins
/ Structure
2014
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Cas1–Cas2 complex formation mediates spacer acquisition during CRISPR–Cas adaptive immunity
Journal Article
Cas1–Cas2 complex formation mediates spacer acquisition during CRISPR–Cas adaptive immunity
2014
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Overview
The CRISPR–Cas system mediates immunity to foreign DNA sequences that are integrated as spacers between repeats in the CRISPR locus. Work from Doudna and colleagues shows that nucleases Cas1 and Cas2 form a stable complex that recognizes the CRISPR leader-repeat sequence, thus determining the site of integration.
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.
Publisher
Nature Publishing Group US,Nature Publishing Group
Subject
/ 45/70
/ 45/77
/ 82/1
/ 82/16
/ 82/80
/ 82/83
/ Clustered Regularly Interspaced Short Palindromic Repeats - physiology
/ CRISPR-Associated Proteins - chemistry
/ CRISPR-Associated Proteins - metabolism
/ CRISPR-Associated Proteins - physiology
/ DNA
/ E coli
/ Endodeoxyribonucleases - chemistry
/ Endodeoxyribonucleases - metabolism
/ Endodeoxyribonucleases - physiology
/ Escherichia coli - immunology
/ Escherichia coli - metabolism
/ Escherichia coli Proteins - chemistry
/ Escherichia coli Proteins - metabolism
/ Escherichia coli Proteins - physiology
/ Genomics
/ Mutation
/ Proteins
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