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Structural biology. Crystal structure of the CRISPR RNA-guided surveillance complex from Escherichia coli.


ABSTRACT: Clustered regularly interspaced short palindromic repeats (CRISPRs) are essential components of RNA-guided adaptive immune systems that protect bacteria and archaea from viruses and plasmids. In Escherichia coli, short CRISPR-derived RNAs (crRNAs) assemble into a 405-kilodalton multisubunit surveillance complex called Cascade (CRISPR-associated complex for antiviral defense). Here we present the 3.24 angstrom resolution x-ray crystal structure of Cascade. Eleven proteins and a 61-nucleotide crRNA assemble into a seahorse-shaped architecture that binds double-stranded DNA targets complementary to the crRNA-guide sequence. Conserved sequences on the 3' and 5' ends of the crRNA are anchored by proteins at opposite ends of the complex, whereas the guide sequence is displayed along a helical assembly of six interwoven subunits that present five-nucleotide segments of the crRNA in pseudo-A-form configuration. The structure of Cascade suggests a mechanism for assembly and provides insights into the mechanisms of target recognition.

SUBMITTER: Jackson RN 

PROVIDER: S-EPMC4188430 | biostudies-literature | 2014 Sep

REPOSITORIES: biostudies-literature

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Structural biology. Crystal structure of the CRISPR RNA-guided surveillance complex from Escherichia coli.

Jackson Ryan N RN   Golden Sarah M SM   van Erp Paul B G PB   Carter Joshua J   Westra Edze R ER   Brouns Stan J J SJ   van der Oost John J   Terwilliger Thomas C TC   Read Randy J RJ   Wiedenheft Blake B  

Science (New York, N.Y.) 20140807 6203


Clustered regularly interspaced short palindromic repeats (CRISPRs) are essential components of RNA-guided adaptive immune systems that protect bacteria and archaea from viruses and plasmids. In Escherichia coli, short CRISPR-derived RNAs (crRNAs) assemble into a 405-kilodalton multisubunit surveillance complex called Cascade (CRISPR-associated complex for antiviral defense). Here we present the 3.24 angstrom resolution x-ray crystal structure of Cascade. Eleven proteins and a 61-nucleotide crRN  ...[more]

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