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DNA Targeting by a Minimal CRISPR RNA-Guided Cascade.

ABSTRACT: Bacteria employ surveillance complexes guided by CRISPR (clustered, regularly interspaced, short palindromic repeats) RNAs (crRNAs) to target foreign nucleic acids for destruction. Although most type I and type III CRISPR systems require four or more distinct proteins to form multi-subunit surveillance complexes, the type I-C systems use just three proteins to achieve crRNA maturation and double-stranded DNA target recognition. We show that each protein plays multiple functional and structural roles: Cas5c cleaves pre-crRNAs and recruits Cas7 to position the RNA guide for DNA binding and unwinding by Cas8c. Cryoelectron microscopy reconstructions of free and DNA-bound forms of the Cascade/I-C surveillance complex reveal conformational changes that enable R-loop formation with distinct positioning of each DNA strand. This streamlined type I-C system explains how CRISPR pathways can evolve compact structures that retain full functionality as RNA-guided DNA capture platforms.

SUBMITTER: Hochstrasser ML

PROVIDER: S-EPMC5111854 | biostudies-literature | 2016 Sep

REPOSITORIES: biostudies-literature

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DNA Targeting by a Minimal CRISPR RNA-Guided Cascade.

Hochstrasser Megan L ML Taylor David W DW Kornfeld Jack E JE Nogales Eva E Doudna Jennifer A JA

Molecular cell 20160901 5

Bacteria employ surveillance complexes guided by CRISPR (clustered, regularly interspaced, short palindromic repeats) RNAs (crRNAs) to target foreign nucleic acids for destruction. Although most type I and type III CRISPR systems require four or more distinct proteins to form multi-subunit surveillance complexes, the type I-C systems use just three proteins to achieve crRNA maturation and double-stranded DNA target recognition. We show that each protein plays multiple functional and structural r ...[more]

PMID: 27588603

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