Structure and Mechanism of the CMR Complex for CRISPR-Mediated Antiviral Immunity
The prokaryotic clusters of regularly interspaced palindromic repeats (CRISPR) system utilizes genomically encoded CRISPR RNA (crRNA), derived from invading viruses and incorporated into ribonucleoprotein complexes with CRISPR-associated (CAS) proteins, to target and degrade viral DNA or RNA on subs...
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| Published in | Molecular cell Vol. 45; no. 3; pp. 303 - 313 |
|---|---|
| Main Authors | , , , , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
United States
Elsevier Inc
10.02.2012
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| Subjects | |
| Online Access | Get full text |
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| Abstract | The prokaryotic clusters of regularly interspaced palindromic repeats (CRISPR) system utilizes genomically encoded CRISPR RNA (crRNA), derived from invading viruses and incorporated into ribonucleoprotein complexes with CRISPR-associated (CAS) proteins, to target and degrade viral DNA or RNA on subsequent infection. RNA is targeted by the CMR complex. In Sulfolobus solfataricus, this complex is composed of seven CAS protein subunits (Cmr1-7) and carries a diverse “payload” of targeting crRNA. The crystal structure of Cmr7 and low-resolution structure of the complex are presented. S. solfataricus CMR cleaves RNA targets in an endonucleolytic reaction at UA dinucleotides. This activity is dependent on the 8 nt repeat-derived 5′ sequence in the crRNA, but not on the presence of a protospacer-associated motif (PAM) in the target. Both target and guide RNAs can be cleaved, although a single molecule of guide RNA can support the degradation of multiple targets.
[Display omitted]
► EM structure of the CMR complex for viral RNA degradation has been determined ► The crRNA content of CMR has been analyzed by deep sequencing ► Target RNA cleavage by CMR is sequence dependent |
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| AbstractList | The prokaryotic clusters of regularly interspaced palindromic repeats (CRISPR) system utilizes genomically encoded CRISPR RNA (crRNA), derived from invading viruses and incorporated into ribonucleoprotein complexes with CRISPR-associated (CAS) proteins, to target and degrade viral DNA or RNA on subsequent infection. RNA is targeted by the CMR complex. In Sulfolobus solfataricus, this complex is composed of seven CAS protein subunits (Cmr1-7) and carries a diverse "payload" of targeting crRNA. The crystal structure of Cmr7 and low-resolution structure of the complex are presented. S. solfataricus CMR cleaves RNA targets in an endonucleolytic reaction at UA dinucleotides. This activity is dependent on the 8 nt repeat-derived 5' sequence in the crRNA, but not on the presence of a protospacer-associated motif (PAM) in the target. Both target and guide RNAs can be cleaved, although a single molecule of guide RNA can support the degradation of multiple targets. The prokaryotic Clusters of Regularly Interspaced Palindromic Repeats (CRISPR) system utilizes genomically-encoded CRISPR RNA (crRNA), derived from invading viruses and incorporated into ribonucleoprotein complexes with CRISPR-associated (CAS) proteins, to target and degrade viral DNA or RNA on subsequent infection. RNA is targeted by the CMR complex. In Sulfolobus solfataricus , this complex is composed of seven CAS protein subunits (Cmr1-7) and carries a diverse “payload” of targeting crRNA. The crystal structure of Cmr7 and low resolution structure of the complex are presented. S. solfataricus CMR cleaves RNA targets in an endonucleolytic reaction at UA dinucleotides. This activity is dependent on the 8-nucleotide repeat-derived 5′ sequence in the crRNA, but not on the presence of a proto-spacer associated motif (PAM) in the target. Both target and guide RNAs can be cleaved, although a single molecule of guide RNA can support the degradation of multiple targets. The prokaryotic clusters of regularly interspaced palindromic repeats (CRISPR) system utilizes genomically encoded CRISPR RNA (crRNA), derived from invading viruses and incorporated into ribonucleoprotein complexes with CRISPR-associated (CAS) proteins, to target and degrade viral DNA or RNA on subsequent infection. RNA is targeted by the CMR complex. In Sulfolobus solfataricus, this complex is composed of seven CAS protein subunits (Cmr1-7) and carries a diverse “payload” of targeting crRNA. The crystal structure of Cmr7 and low-resolution structure of the complex are presented. S. solfataricus CMR cleaves RNA targets in an endonucleolytic reaction at UA dinucleotides. This activity is dependent on the 8 nt repeat-derived 5′ sequence in the crRNA, but not on the presence of a protospacer-associated motif (PAM) in the target. Both target and guide RNAs can be cleaved, although a single molecule of guide RNA can support the degradation of multiple targets. [Display omitted] ► EM structure of the CMR complex for viral RNA degradation has been determined ► The crRNA content of CMR has been analyzed by deep sequencing ► Target RNA cleavage by CMR is sequence dependent The prokaryotic clusters of regularly interspaced palindromic repeats (CRISPR) system utilizes genomically encoded CRISPR RNA (crRNA), derived from invading viruses and incorporated into ribonucleoprotein complexes with CRISPR-associated (CAS) proteins, to target and degrade viral DNA or RNA on subsequent infection. RNA is targeted by the CMR complex. In Sulfolobus solfataricus, this complex is composed of seven CAS protein subunits (Cmr1-7) and carries a diverse "payload" of targeting crRNA. The crystal structure of Cmr7 and low-resolution structure of the complex are presented. S. solfataricus CMR cleaves RNA targets in an endonucleolytic reaction at UA dinucleotides. This activity is dependent on the 8 nt repeat-derived 5' sequence in the crRNA, but not on the presence of a protospacer-associated motif (PAM) in the target. Both target and guide RNAs can be cleaved, although a single molecule of guide RNA can support the degradation of multiple targets. |
| Author | Spagnolo, Laura White, Malcolm F. Reeks, Judith Kerou, Melina Naismith, James H. Albers, Sonja-Verena Graham, Shirley Zhang, Jing Rouillon, Christophe Cannone, Giuseppe Reimann, Julia Brugger, Kim Liu, Huanting |
| AuthorAffiliation | 3 Institute of Structural Molecular Biology and Centre for Science at Extreme Conditions, University of Edinburgh, Edinburgh EH9 3JR, UK 2 EASIH, University of Cambridge, Addenbrookes Hospital, Cambridge CB2 0QQ, UK 1 Biomedical Sciences Research Complex, University of St Andrews, Fife KY16 9ST, UK |
| AuthorAffiliation_xml | – name: 1 Biomedical Sciences Research Complex, University of St Andrews, Fife KY16 9ST, UK – name: 2 EASIH, University of Cambridge, Addenbrookes Hospital, Cambridge CB2 0QQ, UK – name: 3 Institute of Structural Molecular Biology and Centre for Science at Extreme Conditions, University of Edinburgh, Edinburgh EH9 3JR, UK |
| Author_xml | – sequence: 1 givenname: Jing surname: Zhang fullname: Zhang, Jing organization: Biomedical Sciences Research Complex, University of St Andrews, Fife KY16 9ST, UK – sequence: 2 givenname: Christophe surname: Rouillon fullname: Rouillon, Christophe organization: Biomedical Sciences Research Complex, University of St Andrews, Fife KY16 9ST, UK – sequence: 3 givenname: Melina surname: Kerou fullname: Kerou, Melina organization: Biomedical Sciences Research Complex, University of St Andrews, Fife KY16 9ST, UK – sequence: 4 givenname: Judith surname: Reeks fullname: Reeks, Judith organization: Biomedical Sciences Research Complex, University of St Andrews, Fife KY16 9ST, UK – sequence: 5 givenname: Kim surname: Brugger fullname: Brugger, Kim organization: EASIH, University of Cambridge, Addenbrookes Hospital, Cambridge CB2 0QQ, UK – sequence: 6 givenname: Shirley surname: Graham fullname: Graham, Shirley organization: Biomedical Sciences Research Complex, University of St Andrews, Fife KY16 9ST, UK – sequence: 7 givenname: Julia surname: Reimann fullname: Reimann, Julia organization: Archaeal Molecular Biology Group, Max Planck Institute for Terrestrial Microbiology, Karl-von-Frisch-Strasse 10, 35043 Marburg, Germany – sequence: 8 givenname: Giuseppe surname: Cannone fullname: Cannone, Giuseppe organization: Institute of Structural Molecular Biology and Centre for Science at Extreme Conditions, University of Edinburgh, Edinburgh EH9 3JR, UK – sequence: 9 givenname: Huanting surname: Liu fullname: Liu, Huanting organization: Biomedical Sciences Research Complex, University of St Andrews, Fife KY16 9ST, UK – sequence: 10 givenname: Sonja-Verena surname: Albers fullname: Albers, Sonja-Verena organization: Archaeal Molecular Biology Group, Max Planck Institute for Terrestrial Microbiology, Karl-von-Frisch-Strasse 10, 35043 Marburg, Germany – sequence: 11 givenname: James H. surname: Naismith fullname: Naismith, James H. organization: Biomedical Sciences Research Complex, University of St Andrews, Fife KY16 9ST, UK – sequence: 12 givenname: Laura surname: Spagnolo fullname: Spagnolo, Laura email: laura.spagnolo@ed.ac.uk organization: Institute of Structural Molecular Biology and Centre for Science at Extreme Conditions, University of Edinburgh, Edinburgh EH9 3JR, UK – sequence: 13 givenname: Malcolm F. surname: White fullname: White, Malcolm F. email: mfw2@st-andrews.ac.uk organization: Biomedical Sciences Research Complex, University of St Andrews, Fife KY16 9ST, UK |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/22227115$$D View this record in MEDLINE/PubMed |
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| Issue | 3 |
| Language | English |
| License | http://www.elsevier.com/open-access/userlicense/1.0 Copyright © 2012 Elsevier Inc. All rights reserved. https://www.elsevier.com/tdm/userlicense/1.0 |
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| Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 ObjectType-Article-2 ObjectType-Feature-1 In the authors’ opinion Jing Zhang and Christophe Rouillon contributed equally to the work described. |
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| PublicationTitle | Molecular cell |
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| Snippet | The prokaryotic clusters of regularly interspaced palindromic repeats (CRISPR) system utilizes genomically encoded CRISPR RNA (crRNA), derived from invading... The prokaryotic Clusters of Regularly Interspaced Palindromic Repeats (CRISPR) system utilizes genomically-encoded CRISPR RNA (crRNA), derived from invading... |
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| SubjectTerms | Archaeal Proteins - chemistry Archaeal Proteins - isolation & purification Archaeal Viruses - immunology Base Sequence Cellular apoptosis susceptibility protein Crystal structure Crystallography, X-Ray DNA Immunity Infection Inverted Repeat Sequences Macromolecular Substances - chemistry Macromolecular Substances - isolation & purification Microscopy, Electron Models, Molecular Molecular Sequence Data Nucleic Acid Conformation Protein Structure, Quaternary Protein Structure, Tertiary Protein Subunits - chemistry Protein Subunits - isolation & purification Ribonucleoproteins RNA RNA Cleavage RNA, Archaeal - chemistry RNA, Archaeal - genetics RNA, Archaeal - isolation & purification Sulfolobus solfataricus Sulfolobus solfataricus - genetics Sulfolobus solfataricus - immunology Sulfolobus solfataricus - metabolism Sulfolobus solfataricus - virology |
| Title | Structure and Mechanism of the CMR Complex for CRISPR-Mediated Antiviral Immunity |
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