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-Structure paper
Title | DNA Targeting by a Minimal CRISPR RNA-Guided Cascade. |
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Journal, issue, pages | Mol Cell, Vol. 63, Issue 5, Page 840-851, Year 2016 |
Publish date | Sep 1, 2016 |
Authors | Megan L Hochstrasser / David W Taylor / Jack E Kornfeld / Eva Nogales / Jennifer A Doudna / |
PubMed 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 ...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 links | Mol Cell / PubMed:27588603 / PubMed Central |
Methods | EM (single particle) |
Resolution | 6.7 - 8.8 Å |
Structure data | EMDB-8294: EMDB-8295: EMDB-8296: |
Source |
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