CvkR is a MerR-type transcriptional repressor of class 2 type V-K CRISPR-associated transposase systems

Marcus Ziemann, Viktoria Reimann, Yajing Liang, Yue Shi, Honglei Ma, Yuman Xie, Hui Li, Tao Zhu (), Xuefeng Lu () and Wolfgang R. Hess ()
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Marcus Ziemann: University of Freiburg
Viktoria Reimann: University of Freiburg
Yajing Liang: Qingdao Institute of Bioenergy and Bioprocess Technology (QIBEBT), Chinese Academy of Sciences
Yue Shi: Qingdao Institute of Bioenergy and Bioprocess Technology (QIBEBT), Chinese Academy of Sciences
Honglei Ma: Qingdao Institute of Bioenergy and Bioprocess Technology (QIBEBT), Chinese Academy of Sciences
Yuman Xie: Qingdao Institute of Bioenergy and Bioprocess Technology (QIBEBT), Chinese Academy of Sciences
Hui Li: Qingdao Institute of Bioenergy and Bioprocess Technology (QIBEBT), Chinese Academy of Sciences
Tao Zhu: Qingdao Institute of Bioenergy and Bioprocess Technology (QIBEBT), Chinese Academy of Sciences
Xuefeng Lu: Qingdao Institute of Bioenergy and Bioprocess Technology (QIBEBT), Chinese Academy of Sciences
Wolfgang R. Hess: University of Freiburg

Nature Communications, 2023, vol. 14, issue 1, 1-16

Abstract: Abstract Certain CRISPR-Cas elements integrate into Tn7-like transposons, forming CRISPR-associated transposon (CAST) systems. How the activity of these systems is controlled in situ has remained largely unknown. Here we characterize the MerR-type transcriptional regulator Alr3614 that is encoded by one of the CAST (AnCAST) system genes in the genome of cyanobacterium Anabaena sp. PCC 7120. We identify a number of Alr3614 homologs across cyanobacteria and suggest naming these regulators CvkR for Cas V-K repressors. Alr3614/CvkR is translated from leaderless mRNA and represses the AnCAST core modules cas12k and tnsB directly, and indirectly the abundance of the tracr-CRISPR RNA. We identify a widely conserved CvkR binding motif 5’-AnnACATnATGTnnT-3’. Crystal structure of CvkR at 1.6 Å resolution reveals that it comprises distinct dimerization and potential effector-binding domains and that it assembles into a homodimer, representing a discrete structural subfamily of MerR regulators. CvkR repressors are at the core of a widely conserved regulatory mechanism that controls type V-K CAST systems.

Date: 2023
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DOI: 10.1038/s41467-023-36542-9

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