8RC3

DNA bound type IV-A1 CRISPR effector complex from P. oleovorans

Summary for 8RC3

• Entry DOI: 10.2210/pdb8rc3/pdb
• EMDB information: 19046
• Descriptor: CRISPR type AFERR-associated protein Csf2, CRISPR type AFERR-associated protein Csf3, CRISPR type AFERR-associated protein Csf1, ... (8 entities in total)
• Functional Keywords: crispr, crrna, dna binding, type iv crispr-cas, crispri, nuclease deficient, gene regulation
• Biological source: Pseudomonas oleovorans; More
• Total number of polymer chains: 11
• Total formula weight: 320619.50

Authors

Miksys, A.,Cepaite, R.,Malinauskaite, L.,Pausch, P. (deposition date: 2023-12-05, release date: 2024-11-06)

Primary citation

Cepaite, R.,Klein, N.,Miksys, A.,Camara-Wilpert, S.,Ragozius, V.,Benz, F.,Skorupskaite, A.,Becker, H.,Zvejyte, G.,Steube, N.,Hochberg, G.K.A.,Randau, L.,Pinilla-Redondo, R.,Malinauskaite, L.,Pausch, P.
Structural variation of types IV-A1- and IV-A3-mediated CRISPR interference.
Nat Commun, 15:9306-9306, 2024

PubMed Abstract: CRISPR-Cas mediated DNA-interference typically relies on sequence-specific binding and nucleolytic degradation of foreign genetic material. Type IV-A CRISPR-Cas systems diverge from this general mechanism, using a nuclease-independent interference pathway to suppress gene expression for gene regulation and plasmid competition. To understand how the type IV-A system associated effector complex achieves this interference, we determine cryo-EM structures of two evolutionarily distinct type IV-A complexes (types IV-A1 and IV-A3) bound to cognate DNA-targets in the presence and absence of the type IV-A signature DinG effector helicase. The structures reveal how the effector complexes recognize the protospacer adjacent motif and target-strand DNA to form an R-loop structure. Additionally, we reveal differences between types IV-A1 and IV-A3 in DNA interactions and structural motifs that allow for in trans recruitment of DinG. Our study provides a detailed view of type IV-A mediated DNA-interference and presents a structural foundation for engineering type IV-A-based genome editing tools.

PubMed: 39468082
DOI: 10.1038/s41467-024-53778-1

Experimental method

ELECTRON MICROSCOPY (3 Å)