5UZ9

Cryo EM structure of anti-CRISPRs, AcrF1 and AcrF2, bound to type I-F crRNA-guided CRISPR surveillance complex


Experimental Data Snapshot

  • Method: ELECTRON MICROSCOPY
  • Resolution: 3.40 Å
  • Aggregation State: PARTICLE 
  • Reconstruction Method: SINGLE PARTICLE 

wwPDB Validation   3D Report Full Report


This is version 2.2 of the entry. See complete history


Literature

Structure Reveals Mechanisms of Viral Suppressors that Intercept a CRISPR RNA-Guided Surveillance Complex.

Chowdhury, S.Carter, J.Rollins, M.F.Golden, S.M.Jackson, R.N.Hoffmann, C.Nosaka, L.Bondy-Denomy, J.Maxwell, K.L.Davidson, A.R.Fischer, E.R.Lander, G.C.Wiedenheft, B.

(2017) Cell 169: 47-57.e11

  • DOI: https://doi.org/10.1016/j.cell.2017.03.012
  • Primary Citation of Related Structures:  
    5UZ9

  • PubMed Abstract: 

    Genetic conflict between viruses and their hosts drives evolution and genetic innovation. Prokaryotes evolved CRISPR-mediated adaptive immune systems for protection from viral infection, and viruses have evolved diverse anti-CRISPR (Acr) proteins that subvert these immune systems. The adaptive immune system in Pseudomonas aeruginosa (type I-F) relies on a 350 kDa CRISPR RNA (crRNA)-guided surveillance complex (Csy complex) to bind foreign DNA and recruit a trans-acting nuclease for target degradation. Here, we report the cryo-electron microscopy (cryo-EM) structure of the Csy complex bound to two different Acr proteins, AcrF1 and AcrF2, at an average resolution of 3.4 Å. The structure explains the molecular mechanism for immune system suppression, and structure-guided mutations show that the Acr proteins bind to residues essential for crRNA-mediated detection of DNA. Collectively, these data provide a snapshot of an ongoing molecular arms race between viral suppressors and the immune system they target.


  • Organizational Affiliation

    Department of Integrative Structural and Computational Biology, Scripps Research Institute, La Jolla, CA 92037, USA.


Macromolecules

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Entity ID: 1
MoleculeChains Sequence LengthOrganismDetailsImage
CRISPR-associated protein Csy1434Pseudomonas aeruginosa UCBPP-PA14Mutation(s): 0 
Gene Names: csy1PA14_33330
UniProt
Find proteins for Q02ML9 (Pseudomonas aeruginosa (strain UCBPP-PA14))
Explore Q02ML9 
Go to UniProtKB:  Q02ML9
Entity Groups  
Sequence Clusters30% Identity50% Identity70% Identity90% Identity95% Identity100% Identity
UniProt GroupQ02ML9
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Entity ID: 2
MoleculeChains Sequence LengthOrganismDetailsImage
CRISPR-associated protein Csy2327Pseudomonas aeruginosa UCBPP-PA14Mutation(s): 0 
Gene Names: csy2PA14_33320
UniProt
Find proteins for Q02MM0 (Pseudomonas aeruginosa (strain UCBPP-PA14))
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Go to UniProtKB:  Q02MM0
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UniProt GroupQ02MM0
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Entity ID: 3
MoleculeChains Sequence LengthOrganismDetailsImage
CRISPR-associated protein Csy3
C, D, E, F, G
C, D, E, F, G, H
341Pseudomonas aeruginosa UCBPP-PA14Mutation(s): 0 
Gene Names: csy3csy1-3PA14_33310
UniProt
Find proteins for Q02MM1 (Pseudomonas aeruginosa (strain UCBPP-PA14))
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Go to UniProtKB:  Q02MM1
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UniProt GroupQ02MM1
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Entity ID: 4
MoleculeChains Sequence LengthOrganismDetailsImage
Anti-CRISPR protein Acr30-35
I, J
77Pseudomonas phage JBD30Mutation(s): 0 
Gene Names: JBD30_035
UniProt
Find proteins for L7P7M1 (Pseudomonas phage JBD30)
Explore L7P7M1 
Go to UniProtKB:  L7P7M1
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UniProt GroupL7P7M1
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Entity ID: 5
MoleculeChains Sequence LengthOrganismDetailsImage
Anti-CRISPR protein 3096Casadabanvirus D3112Mutation(s): 0 
Gene Names: orf30
UniProt
Find proteins for Q6TM72 (Pseudomonas phage D3112)
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UniProt GroupQ6TM72
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Entity ID: 6
MoleculeChains Sequence LengthOrganismDetailsImage
CRISPR-associated endonuclease Cas6/Csy4189Pseudomonas aeruginosa UCBPP-PA14Mutation(s): 0 
Gene Names: cas6fcsy4PA14_33300
EC: 3.1
UniProt
Find proteins for Q02MM2 (Pseudomonas aeruginosa (strain UCBPP-PA14))
Explore Q02MM2 
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UniProt GroupQ02MM2
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Entity ID: 7
MoleculeChains LengthOrganismImage
CRISPR RNA (60-MER)60Pseudomonas aeruginosa UCBPP-PA14
Sequence Annotations
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Experimental Data & Validation

Experimental Data

  • Method: ELECTRON MICROSCOPY
  • Resolution: 3.40 Å
  • Aggregation State: PARTICLE 
  • Reconstruction Method: SINGLE PARTICLE 
EM Software:
TaskSoftware PackageVersion
RECONSTRUCTIONRELION1.4
MODEL REFINEMENTPHENIX1.11.1
MODEL REFINEMENTPHENIX1.11.1
MODEL REFINEMENTPHENIX1.11.1

Structure Validation

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Entry History & Funding Information

Deposition Data


Funding OrganizationLocationGrant Number
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)United StatesP20GM103500, P30GM110732-03, R01GM110270, R01GM108888, P20GM103474, F32GM108436, DP2EB020402, DP5OD021344
National Science Foundation (NSF, United States)United StatesEPS-110134
Canadian Institutes of Health Research (CIHR)CanadaMOP-130482, MOP-136845

Revision History  (Full details and data files)

  • Version 1.0: 2017-04-26
    Type: Initial release
  • Version 1.1: 2017-09-27
    Changes: Author supporting evidence, Data collection
  • Version 1.2: 2018-07-18
    Changes: Data collection, Experimental preparation
  • Version 2.0: 2019-11-06
    Changes: Atomic model, Data collection, Other
  • Version 2.1: 2019-11-27
    Changes: Author supporting evidence
  • Version 2.2: 2024-03-13
    Changes: Data collection, Database references