Crystal structure of the Csm1 subunit of the Csm complex and its single-stranded DNA-specific nuclease activity

Structure
Tae-Yang JungEuijeon Woo

Abstract

The CRISPR-Cas system is the RNA-guided immune defense mechanism in bacteria and archaea. Csm1 belongs to the Cas10 family, which is the common signature protein of the type III system. Csm1 is the largest subunit of the Csm interference complex in the type III-A subtype, which targets foreign DNA or RNA. Here, we report crystallographic and biochemical analyses of Thermococcus onnurineus Csm1, revealing a five-domain organization and single-stranded DNA (ssDNA)-specific nuclease activity associated with the N-terminal HD domain. This domain folds into permuted secondary structures in comparison with the HD domain of Cas3 and contains all the catalytically important residues. It exhibited both endo- and exonuclease activities in an Ni(2+) or Mn(2+)-dependent manner. The narrow width of the active-site cleft appears to restrict the substrate specificity to ssDNA and thus to prevent Csm1 from cleaving double-stranded chromosomal DNA. These data suggest that Csm1 may function in DNA interference by the Csm effector complex.

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Citations

Jan 24, 2016·Nucleic Acids Research·Jing ZhangMalcolm F White
Sep 29, 2015·Nature Reviews. Microbiology·Kira S MakarovaEugene V Koonin
Oct 16, 2015·Nucleic Acids Research·Yingjun LiQunxin She
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Jul 3, 2018·ELife·Christophe RouillonMalcolm F White
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Apr 26, 2019·The CRISPR Journal·Kira S MakarovaEugene V Koonin
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Jun 8, 2021·Frontiers in Microbiology·Hannah N TaylorRyan N Jackson
Feb 6, 2016·Genes & Development·Joshua R ElmoreMichael P Terns
Feb 6, 2016·Genes & Development·Michael A EstrellaScott Bailey

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