title: Two Distant Catalytic Sites Are Responsible for C2c2 RNase Activities.
abstract: C2c2, the effector of type VI CRISPR-Cas systems, has two RNase activities-one for cutting its RNA target and the other for processing the CRISPR RNA (crRNA). Here, we report the structures of Leptotrichia shahii C2c2 in its crRNA-free and crRNA-bound states. While C2c2 has a bilobed structure reminiscent of all other Class 2 effectors, it also exhibits different structural characteristics. It contains the REC lobe with a Helical-1 domain and the NUC lobe with two HEPN domains. The two RNase catalytic pockets responsible for cleaving pre-crRNA and target RNA are independently located on Helical-1 and HEPN domains, respectively. crRNA binding induces significant conformational changes that are likely to stabilize crRNA binding and facilitate target RNA recognition. These structures provide important insights into the molecular mechanism of dual RNase activities of C2c2 and establish a framework for its future engineering as a RNA editing tool.
authors.affiliation: Key Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Liu
authors.affiliation: Key Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Xueyan
authors.initials: X
authors.lastname: Li
authors.affiliation: Key Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Jiuyu
authors.initials: J
authors.lastname: Wang
authors.affiliation: Key Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Min
authors.initials: M
authors.lastname: Wang
authors.affiliation: Key Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Chen
authors.affiliation: Key Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Maolu
authors.initials: M
authors.lastname: Yin
authors.affiliation: Key Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Jiazhi
authors.initials: J
authors.lastname: Li
authors.affiliation: Key Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Sheng
authors.affiliation: Key Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China; Collaborative Innovation Center of Genetics and Development, Shanghai 200438, China. Electronic address: ylwang@ibp.ac.cn.
authors.forename: Yanli
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D064113:CRISPR-Cas Systems; D020134:Catalytic Domain; D045183:Leptotrichia / Q000737:chemistry* / Q000145:classification / Q000201:enzymology* / Q000378:metabolism; D008958:Models, Molecular; D016296:Mutagenesis; D012323:RNA Processing, Post-Transcriptional; D012329:RNA, Bacterial / Q000737:chemistry; D022661:RNA, Untranslated / Q000737:chemistry; D012260:Ribonucleases / Q000737:chemistry*; D016415:Sequence Alignment
chemical_list: D012329:RNA, Bacterial; D022661:RNA, Untranslated; D012260:Ribonucleases
keywords: C2c2; CRISPR-Cas; HEPN domain; RNA-guided RNA cleavage; endonuclease; precursor crRNA