You are here

Bipartite recognition of target RNAs activates DNA cleavage by the Type III-B CRISPR-Cas system.

Title: Bipartite recognition of target RNAs activates DNA cleavage by the Type III-B CRISPR-Cas system.
3 views
0 downloads
Name(s): Elmore, Joshua R, author
Sheppard, Nolan F, author
Ramia, Nancy, author
Deighan, Trace, author
Li, Hong, author
Terns, Rebecca M, author
Terns, Michael P, author
Type of Resource: text
Genre: Journal Article
Text
Date Issued: 2016-02-15
Physical Form: computer
online resource
Extent: 1 online resource
Language(s): English
Abstract/Description: CRISPR-Cas systems eliminate nucleic acid invaders in bacteria and archaea. The effector complex of the Type III-B Cmr system cleaves invader RNAs recognized by the CRISPR RNA (crRNA ) of the complex. Here we show that invader RNAs also activate the Cmr complex to cleave DNA. As has been observed for other Type III systems, Cmr eliminates plasmid invaders in Pyrococcus furiosus by a mechanism that depends on transcription of the crRNA target sequence within the plasmid. Notably, we found that the target RNA per se induces DNA cleavage by the Cmr complex in vitro. DNA cleavage activity does not depend on cleavage of the target RNA but notably does require the presence of a short sequence adjacent to the target sequence within the activating target RNA (rPAM [RNA protospacer-adjacent motif]). The activated complex does not require a target sequence (or a PAM) in the DNA substrate. Plasmid elimination by the P. furiosus Cmr system also does not require the Csx1 (CRISPR-associated Rossman fold [CARF] superfamily) protein. Plasmid silencing depends on the HD nuclease and Palm domains of the Cmr2 (Cas10 superfamily) protein. The results establish the Cmr complex as a novel DNA nuclease activated by invader RNAs containing a crRNA target sequence and a rPAM.
Identifier: FSU_pmch_26848045 (IID), 10.1101/gad.272153.115 (DOI), PMC4762429 (PMCID), 26848045 (RID), 26848045 (EID), gad.272153.115 (PII)
Keywords: CRISPR–Cas, Cmr, DNA nuclease, PAM, RNAi
Grant Number: R01 GM054682, R35 GM118160, R01 GM54682
Publication Note: This NIH-funded author manuscript originally appeared in PubMed Central at https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4762429.
Subject(s): Bacterial Proteins/genetics
Bacterial Proteins/metabolism
CRISPR-Cas Systems/physiology
DNA Cleavage
Deoxyribonucleases/metabolism
Enzyme Activation
Gene Silencing/physiology
Plasmids/genetics
Protein Binding
Pyrococcus furiosus/physiology
RNA/metabolism
Persistent Link to This Record: http://purl.flvc.org/fsu/fd/FSU_pmch_26848045
Owner Institution: FSU
Is Part Of: Genes & development.
1549-5477
Issue: iss. 4, vol. 30

Choose the citation style.
Elmore, J. R., Sheppard, N. F., Ramia, N., Deighan, T., Li, H., Terns, R. M., & Terns, M. P. (2016). Bipartite recognition of target RNAs activates DNA cleavage by the Type III-B CRISPR-Cas system. Genes & Development. Retrieved from http://purl.flvc.org/fsu/fd/FSU_pmch_26848045