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Structural variation of types IV-A1- and IV-A3-mediated CRISPR interference

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 determined cryo-EM structures of two evolutionarily distinct type IV-A complexes (types IV-38 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.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Pseudomonas Oleovorans

SUBMITTER: Rimvydė Čepaitė

LAB HEAD: Patrick Pausch

PROVIDER: PXD056399 | Pride | 2024-10-08

REPOSITORIES: Pride

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			Action	DRS
	B1_ion.tsv	Tabular
	B1_peptide.tsv	Tabular
	B1_protein.tsv	Tabular
	B1_psm.tsv	Tabular
	B2_ion.tsv	Tabular

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