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TitleMolecular mechanism of active Cas7-11 in processing CRISPR RNA and interfering target RNA.
Journal, issue, pagesElife, Vol. 11, Year 2022
Publish dateOct 3, 2022
AuthorsHemant N Goswami / Jay Rai / Anuska Das / Hong Li /
PubMed AbstractCas7-11 is a Type III-E CRISPR Cas effector that confers programmable RNA cleavage and has potential applications in RNA interference. Cas7-11 encodes a single polypeptide containing four Cas7- and ...Cas7-11 is a Type III-E CRISPR Cas effector that confers programmable RNA cleavage and has potential applications in RNA interference. Cas7-11 encodes a single polypeptide containing four Cas7- and one Cas11-like segments that obscures the distinction between the multi-subunit Class 1 and the single-subunit Class-2 CRISPR Cas systems. We report a cryo-EM (cryo-electron microscopy) structure of the active Cas7-11 from (DiCas7-11) that reveals the molecular basis for RNA processing and interference activities. DiCas7-11 arranges its Cas7- and Cas11-like domains in an extended form that resembles the backbone made up by four Cas7 and one Cas11 subunits in the multi-subunit enzymes. Unlike the multi-subunit enzymes, however, the backbone of DiCas7-11 contains evolutionarily different Cas7 and Cas11 domains, giving rise to their unique functionality. The first Cas7-like domain nearly engulfs the last 15 direct repeat nucleotides in processing and recognition of the CRISPR RNA, and its free-standing fragment retains most of the activity. Both the second and the third Cas7-like domains mediate target RNA cleavage in a metal-dependent manner. The structure and mutational data indicate that the long variable insertion to the fourth Cas7 domain has little impact on RNA processing or targeting, suggesting the possibility for engineering a compact and programmable RNA interference tool.
External linksElife / PubMed:36190192 / PubMed Central
MethodsEM (single particle)
Resolution2.82 Å
Structure data

27138

8d1v

EMDB-27138, PDB-8d1v:
Cryo-EM structure of guide RNA and target RNA bound Cas7-11
Method: EM (single particle) / Resolution: 2.82 Å

Chemicals

ChemComp-ZN:
Unknown entry

Source
  • desulfonema ishimotonii (bacteria)
KeywordsRNA BINDING PROTEIN/RNA / Crispr / type III-E Cas7-11 / RNA BINDING PROTEIN / RNA BINDING PROTEIN-RNA complex

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