title: Staphylococcus epidermidis Csm1 is a 3'-5' exonuclease.
abstract: Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) offer an adaptive immune system that protects bacteria and archaea from nucleic acid invaders through an RNA-mediated nucleic acid cleavage mechanism. Our knowledge of nucleic acid cleavage mechanisms is limited to three examples of widely different ribonucleoprotein particles that target either DNA or RNA. Staphylococcus epidermidis belongs to the Type III-A CRISPR system and has been shown to interfere with invading DNA in vivo. The Type III-A CRISPR system is characterized by the presence of Csm1, a member of Cas10 family of proteins, that has a permuted histidine-aspartate domain and a nucleotidyl cyclase-like domain, both of which contain sequence features characteristic of nucleases. In this work, we show in vitro that a recombinant S. epidermidis Csm1 cleaves single-stranded DNA and RNA exonucleolytically in the 3'-5' direction. We further showed that both cleavage activities are divalent-metal-dependent and reside in the GGDD motif of the cyclase-like domain. Our data suggest that Csm1 may work in the context of an effector complex to degrade invading DNA and participate in CRISPR RNA maturation.
authors.affiliation: Institute of Molecular Biophysics, Florida State University, Tallahassee, FL 32306, USA and Department of Chemistry and Biochemistry, Florida State University, Tallahassee, FL 32306, USA.
authors.forename: Nancy F
authors.initials: NF
authors.lastname: Ramia
authors.forename: Li
authors.initials: L
authors.lastname: Tang
authors.forename: Alexis I
authors.initials: AI
authors.lastname: Cocozaki
authors.forename: Hong
authors.initials: H
authors.lastname: Li
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D020134:Catalytic Domain; D002413:Cations, Divalent; D004277:DNA, Single-Stranded / Q000378:metabolism; D005092:Exonucleases / Q000737:chemistry* / Q000378:metabolism*; D008670:Metals / Q000737:chemistry; D012313:RNA / Q000378:metabolism; D013212:Staphylococcus epidermidis / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D002413:Cations, Divalent; D004277:DNA, Single-Stranded; D008670:Metals; D012313:RNA; D005092:Exonucleases