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TitleDNA Targeting by a Minimal CRISPR RNA-Guided Cascade.
Journal, issue, pagesMol Cell, Vol. 63, Issue 5, Page 840-851, Year 2016
Publish dateSep 1, 2016
AuthorsMegan L Hochstrasser / David W Taylor / Jack E Kornfeld / Eva Nogales / Jennifer A Doudna /
PubMed AbstractBacteria employ surveillance complexes guided by CRISPR (clustered, regularly interspaced, short palindromic repeats) RNAs (crRNAs) to target foreign nucleic acids for destruction. Although most ...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.
External linksMol Cell / PubMed:27588603 / PubMed Central
MethodsEM (single particle)
Resolution6.7 - 8.8 Å
Structure data

EMDB-8294:
Structure of 2x duplex target DNA-bound Cascade/I-C
Method: EM (single particle) / Resolution: 6.7 Å

EMDB-8295:
Structure of 2x duplex DNA target-bound Cascade/I-C
Method: EM (single particle) / Resolution: 8.8 Å

EMDB-8296:
Structure of 1x duplex DNA target-bound Cascade/I-C
Method: EM (single particle) / Resolution: 7.6 Å

Source
  • Desulfovibrio vulgaris (bacteria)

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