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. 2020 Apr 29;48(11):6149–6156. doi: 10.1093/nar/gkaa298

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© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — The atypical type III CRISPR system of M. piezophila. (A) CRISPR locus of M. piezophila (NCBI accession NC_016751) includes genes encoding a complete type III-B (Cmr1–6, Cas6) gene cluster, adaptation proteins Cas1 and Cas2, argonaute, and two uncharacterised CARF-domain proteins: CARF-RelE and Csx1-Crn2. (B) Cartoon of the Csx1-Crn2 protein highlighting the N-terminal Csx1 ribonuclease and C-terminal Crn2 ring nuclease components. (C) Structural model of Csx1–Crn2. The active site residues of the HEPN ribonuclease (R397, H402) and ring nuclease (H495) are shown in spheres. Domain colouring is as for part B. Although the structures of the Csx1 and Crn2 domains are predicted with high confidence, their relative orientation is not known and the model shown here represents just one possible conformation. (D) SDS-PAGE analysis of purified recombinant Csx1Crn2 wild-type and variants.