› 5h9f

X-ray diffraction
2.45Å resolution

Crystal structure of E. coli Cascade bound to a PAM-containing dsDNA target at 2.45 angstrom resolution.

Released:
DOI: 10.2210/pdb5h9f/pdb
PDB 5h9f coloured by chain and viewed from the front.
PDB 5h9f coloured by chain and viewed from the side.
PDB 5h9f coloured by chain and viewed from the top.
Source organism: Escherichia coli K-12
Primary publication:
Structural basis for promiscuous PAM recognition in type I-E Cascade from E. coli.
Hayes RP, Xiao Y, Ding F, van Erp PB, Rajashankar K, Bailey S, Wiedenheft B, Ke A
Nature 530 499-503 (2016)
PMID: 26863189

Function and Biology Details

Biochemical function:
Biological process:
Cellular component:
Sequence domains:

Structure analysis Details

Assembly composition:
hetero tetradecamer (preferred)
Entry contents:
5 distinct polypeptide molecules
2 distinct DNA molecules
1 distinct RNA molecule
Macromolecules (8 distinct):
CRISPR system Cascade subunit CasA Chain: A
Molecule details ›
Chain: A
Length: 502 amino acids
Theoretical weight: 55.96 KDa
Source organism: Escherichia coli K-12
Expression system: Escherichia coli
UniProt:
  • Canonical: Q46901 (Residues: 1-502; Coverage: 100%)
Gene names: JW2730, b2760, casA, cse1, ygcL
Sequence domains: CRISPR-associated protein Cse1 (CRISPR_cse1)
CRISPR system Cascade subunit CasB Chains: B, C
Molecule details ›
Chains: B, C
Length: 165 amino acids
Theoretical weight: 19.23 KDa
Source organism: Escherichia coli K-12
Expression system: Escherichia coli
UniProt:
  • Canonical: P76632 (Residues: 1-160; Coverage: 100%)
Gene names: JW2729, b2759, casB, cse2, ygcK
Sequence domains: CRISPR-associated protein Cse2 (CRISPR_cse2)
Structure domains: CRISPR-associated protein Cse2
CRISPR system Cascade subunit CasC Chains: D, E, F, G, H, I
Molecule details ›
Chains: D, E, F, G, H, I
Length: 363 amino acids
Theoretical weight: 40.07 KDa
Source organism: Escherichia coli K-12
Expression system: Escherichia coli
UniProt:
  • Canonical: Q46899 (Residues: 1-363; Coverage: 100%)
Gene names: JW2728, b2758, cas4, casC, cse4, ygcJ
Sequence domains: CT1975-like protein
CRISPR system Cascade subunit CasD Chain: J
Molecule details ›
Chain: J
Length: 224 amino acids
Theoretical weight: 25.24 KDa
Source organism: Escherichia coli K-12
Expression system: Escherichia coli
UniProt:
  • Canonical: Q46898 (Residues: 1-224; Coverage: 100%)
Gene names: JW5844, b2757, cas5, casD, ygcI
Sequence domains: CRISPR-associated protein (Cas_Cas5)
Structure domains: Alpha-Beta Plaits
CRISPR system Cascade subunit CasE Chain: K
Molecule details ›
Chain: K
Length: 199 amino acids
Theoretical weight: 22.32 KDa
Source organism: Escherichia coli K-12
Expression system: Escherichia coli
UniProt:
  • Canonical: Q46897 (Residues: 1-199; Coverage: 100%)
Gene names: JW2726, b2756, cas6e, casE, ygcH
Sequence domains: CRISPR associated protein
Structure domains: Crispr-associated protein; domain 2
DNA (28-MER) Non-target Chain: M
Molecule details ›
Chain: M
Length: 28 nucleotides
Theoretical weight: 8.6 KDa
DNA (50-MER) Target Chain: N
Molecule details ›
Chain: N
Length: 50 nucleotides
Theoretical weight: 15.39 KDa
crRNA (61-MER) Chain: L
Molecule details ›
Chain: L
Length: 61 nucleotides
Theoretical weight: 19.68 KDa

Ligands and Environments

1 bound ligand:
Ligand ZN 1 x ZN
1 modified residue:
Ligand 23G 1 x 23G

Experiments and Validation Details

Entry percentile scores
X-ray source: APS BEAMLINE 24-ID-C
Unit cell:
a: 92.979Å b: 150.059Å c: 400.546Å
α: 90° β: 90° γ: 90°
R-values:
R R work R free
0.206 0.206 0.233
Expression system: Escherichia coli

Quick links

Citations

18 review citations

Chemistry of Class 1 CRISPR-Cas effectors: Binding, editing, and regulation.
Liu et al. (2020)
17 more

1 mention without citation

CRISPR-Mediated Epigenome Editing.
Enríquez P. (2016)