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TitleTarget RNA-guided protease activity in type III-E CRISPR-Cas system.
Journal, issue, pagesNucleic Acids Res, Vol. 50, Issue 22, Page 12913-12923, Year 2022
Publish dateDec 9, 2022
AuthorsXiaoshen Wang / Guimei Yu / Yanan Wen / Qiyin An / Xuzichao Li / Fumeng Liao / Chengwei Lian / Kai Zhang / Hang Yin / Yong Wei / Zengqin Deng / Heng Zhang /
PubMed AbstractThe type III-E CRISPR-Cas systems are newly identified adaptive immune systems in prokaryotes that use a single Cas7-11 protein to specifically cleave target RNA. Cas7-11 could associate with Csx29, ...The type III-E CRISPR-Cas systems are newly identified adaptive immune systems in prokaryotes that use a single Cas7-11 protein to specifically cleave target RNA. Cas7-11 could associate with Csx29, a putative caspase-like protein encoded by the gene frequently found in the type III-E loci, suggesting a functional linkage between the RNase and protease activities in type III-E systems. Here, we demonstrated that target RNA recognition would stimulate the proteolytic activity of Csx29, and protein Csx30 is the endogenous substrate. More interestingly, while the cognate target RNA recognition would activate Csx29, non-cognate target RNA with the complementary 3' anti-tag sequence inhibits the enzymatic activity. Csx30 could bind to the sigma factor RpoE, which may initiate the stress response after proteolytic cleavage. Combined with biochemical and structural studies, we have elucidated the mechanisms underlying the target RNA-guided proteolytic activity of Csx29. Our work will guide further developments leveraging this simple RNA targeting system for RNA and protein-related applications.
External linksNucleic Acids Res / PubMed:36484100 / PubMed Central
MethodsEM (single particle)
Resolution2.8 - 3.1 Å
Structure data

34158

8gna

EMDB-34158, PDB-8gna:
Structure of the SbCas7-11-crRNA-NTR complex
Method: EM (single particle) / Resolution: 2.8 Å

34270

8gu6

EMDB-34270, PDB-8gu6:
Structure of the SbCas7-11-crRNA-NTR-Csx29 complex
Method: EM (single particle) / Resolution: 3.1 Å

Chemicals

ChemComp-ZN:
Unknown entry

Source
  • candidatus scalindua brodae (bacteria)
KeywordsRNA BINDING PROTEIN / Complex / RNA-binding

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