title: Active site plasticity enables metal-dependent tuning of Cas5d nuclease activity in CRISPR-Cas type I-C system.
abstract: Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) in association with CRISPR-associated (Cas) proteins constitutes a formidable defense system against mobile genetic elements in prokaryotes. In type I-C, the ribonucleoprotein surveillance complex comprises only three Cas proteins, namely, Cas5d, Csd1 and Csd2. Unlike type I-E that uses Cse3/CasE for metal-independent CRISPR RNA maturation, type I-C that lacks this deputes Cas5d to process the pre-crRNA. Here, we report the promiscuous DNase activity of Cas5d in presence of divalent metals. Remarkably, the active site that renders RNA hydrolysis may be tuned by metal to act on DNA substrates too. Further, the realization that Csd1 is a fusion of its functional homolog Cse1/CasA and Cse2/CasB forecasts that the stoichiometry of the constituents of the surveillance complex in type I-C may differ from type I-E. Although Csd2 seems to be inert, Csd1 too exhibits RNase and metal-dependent DNase activity. Thus, in addition to their proposed functions, the DNase activity of Cas5d and Csd1 may also enable them to be co-opted in adaptation and interference stages of CRISPR immunity wherein interaction with DNA substrates is involved.
authors.affiliation: Department of Biotechnology, Indian Institute of Technology Guwahati, Guwahati 781039, India.
authors.forename: Ankita
authors.initials: A
authors.lastname: Punetha
authors.forename: Raveendran
authors.initials: R
authors.lastname: Sivathanu
authors.forename: Baskaran
authors.initials: B
authors.lastname: Anand
mesh_terms: D064130:CRISPR-Associated Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D064113:CRISPR-Cas Systems; D020134:Catalytic Domain; D002413:Cations, Divalent; D004706:Endodeoxyribonucleases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008670:Metals / Q000737:chemistry / Q000378:metabolism*
chemical_list: D064130:CRISPR-Associated Proteins; D002413:Cations, Divalent; D008670:Metals; D004706:Endodeoxyribonucleases