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Structure paper

TitleCryo-EM Structures Reveal Mechanism and Inhibition of DNA Targeting by a CRISPR-Cas Surveillance Complex.
Journal, issue, pagesCell, Vol. 171, Issue 2, Page 414-426.e12, Year 2017
Publish dateOct 5, 2017
AuthorsTai Wei Guo / Alberto Bartesaghi / Hui Yang / Veronica Falconieri / Prashant Rao / Alan Merk / Edward T Eng / Ashleigh M Raczkowski / Tara Fox / Lesley A Earl / Dinshaw J Patel / Sriram Subramaniam /
PubMed AbstractProkaryotic cells possess CRISPR-mediated adaptive immune systems that protect them from foreign genetic elements, such as invading viruses. A central element of this immune system is an RNA-guided ...Prokaryotic cells possess CRISPR-mediated adaptive immune systems that protect them from foreign genetic elements, such as invading viruses. A central element of this immune system is an RNA-guided surveillance complex capable of targeting non-self DNA or RNA for degradation in a sequence- and site-specific manner analogous to RNA interference. Although the complexes display considerable diversity in their composition and architecture, many basic mechanisms underlying target recognition and cleavage are highly conserved. Using cryoelectron microscopy (cryo-EM), we show that the binding of target double-stranded DNA (dsDNA) to a type I-F CRISPR system yersinia (Csy) surveillance complex leads to large quaternary and tertiary structural changes in the complex that are likely necessary in the pathway leading to target dsDNA degradation by a trans-acting helicase-nuclease. Comparison of the structure of the surveillance complex before and after dsDNA binding, or in complex with three virally encoded anti-CRISPR suppressors that inhibit dsDNA binding, reveals mechanistic details underlying target recognition and inhibition.
External linksCell / PubMed:28985564 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution2.265 - 3.6 Å
Structure data

7048

6b44

EMDB-7048, PDB-6b44:
Cryo-EM structure of Type I-F CRISPR crRNA-guided Csy surveillance complex with bound target dsDNA
Method: EM (single particle) / Resolution: 2.9 Å

7049

6b45

EMDB-7049, PDB-6b45:
Cryo-EM structure of Type I-F CRISPR crRNA-guided Csy surveillance complex
Method: EM (single particle) / Resolution: 3.5 Å

7050

6b46

EMDB-7050, PDB-6b46:
Cryo-EM structure of Type I-F CRISPR crRNA-guided Csy surveillance complex with bound anti-CRISPR protein AcrF1
Method: EM (single particle) / Resolution: 3.1 Å

7051

6b47

EMDB-7051, PDB-6b47:
Cryo-EM structure of Type I-F CRISPR crRNA-guided Csy surveillance complex with bound anti-CRISPR protein AcrF2
Method: EM (single particle) / Resolution: 3.2 Å

7052

6b48

EMDB-7052, PDB-6b48:
Cryo-EM structure of Type I-F CRISPR crRNA-guided Csy surveillance complex with bound anti-CRISPR protein AcrF10
Method: EM (single particle) / Resolution: 3.6 Å

PDB-6anv:
Crystal structure of anti-CRISPR protein AcrF1
Method: X-RAY DIFFRACTION / Resolution: 2.265 Å

PDB-6anw:
Crystal structure of anti-CRISPR protein AcrF10
Method: X-RAY DIFFRACTION / Resolution: 2.486 Å

Chemicals

ChemComp-MES:
2-(N-MORPHOLINO)-ETHANESULFONIC ACID / pH buffer*YM / MES (buffer)

ChemComp-PGE:
TRIETHYLENE GLYCOL / Polyethylene glycol

ChemComp-EDO:
1,2-ETHANEDIOL / Ethylene glycol

ChemComp-PEG:
DI(HYDROXYETHYL)ETHER / Diethylene glycol

ChemComp-HOH:
WATER / Water

Source
  • pseudomonas aeruginosa (bacteria)
  • pseudomonas aeruginosa (strain ucbpp-pa14) (bacteria)
  • synthetic construct (others)
  • pseudomonas phage jbd30 (virus)
  • pseudomonas phage d3112 (virus)
  • shewanella xiamenensis (bacteria)
  • escherichia coli o157:h7 (bacteria)
KeywordsIMMUNE SYSTEM / Type I-F CRISPR-Cas system: Csy Cascade: Structure: anti-CRISPR protein: Inhibition of Csy complex: Genome editing tool / IMMUNE SYSTEM/RNA/DNA / CRISPR-Cas / IMMUNE SYSTEM-RNA-DNA complex / IMMUNE SYSTEM / RNA / IMMUNE SYSTEM - RNA complex / IMMUNE SYSTEM/HYDROLASE/RNA / IMMUNE SYSTEM-HYDROLASE-RNA complex

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