Dual nuclease activity of a Cas2 protein in <scp>CRISPR</scp>–Cas subtype I‐B of <i>Leptospira interrogans</i>

Dixit, Bhuvan; Ghosh, Karukriti Kaushik; Fernandes, Gary; Kumar, Pankaj; Gogoi, Prerana; Kumar, Manish

doi:10.1002/1873-3468.12124

Cited by 27 publications

(31 citation statements)

References 51 publications

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“…For instance, it has been demonstrated that Cas2 of Legionella pneumophila displays nuclease activity, which promotes the intracellular infection of amoebal host cells (31). Since L. interrogans subtype I-C Cas2 exhibited nuclease activity against dsDNA and Cas2 proteins in CRISPR-Cas subtype I-B possess nuclease activity as well (32), we speculated that the extra subtype I-C hit to leptospira plasmid hit to leptospira phage hit to leptospira plasmid and other virus hit to leptospira phage and other plasmid hit to other plasmid hit to other phage hit to both phage and plasmid hit to other virus 1 2 3 4 5 6 1 2 3 4 5 6 7 8 9 10 1 1 12 13 14 15 16 17 18 19 20 21 22 23 24 CRISPR-Cas system may be responsible for the difficulty in transforming the DNA into Leptospira for genetic manipulation.…”

Section: Discussionmentioning

confidence: 99%

Characterization of CRISPR‐Cas systems in Leptospira reveals potential application of CRISPR in genotyping of Leptospira interrogans

Gao

Che

et al. 2019

APMIS

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Leptospirosis is a zoonotic disease caused by pathogenic Leptospira. However, understanding of the pathogenic mechanism of Leptospira is still elusive due to the limited number of genetic tools available for this microorganism. Currently, the reason for the genetic inaccessibility of Leptospira is still unknown. It is well known that as an acquired immunity of bacteria, Clustered Regularly Interspaced Short Palindromic Repeat‐CRISPR‐associated gene (CRISPR‐Cas) systems can help bacteria against invading mobile genetic elements. In this study, the occurrence and diversity of CRISPR‐Cas systems in 41 genomes of Leptospira strains were investigated. Three subtypes (subtype I‐B, subtype I‐C and subtype I‐E) of CRISPR‐Cas systems were identified in both pathogenic and intermediate Leptospira species but not in saprophytic species. Noteworthy, the majority of pathogenic species harbor two different types of CRISPR‐Cas systems (subtype I‐B and subtype I‐E). Furthermore, Cas2 protein of subtype I‐C in L. interrogans exhibited a metal‐dependent DNase activity in a nonspecific manner. CRISPR spacers in subtype I‐B are highly conserved within the same serovars and hypervariable across different serovars of L. interrogans. Based on the subtype I‐B CRISPR arrays, the serotypes of different L. interrogans strains were easily identified. Investigation of the origin of CRISPR spacers showed that 192 spacers (23.5%) matched to mobile genetic elements, indicating CRISPR‐Cas systems may play an important role in the defense of foreign invading DNA.

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Section: Discussionmentioning

confidence: 99%

Characterization of CRISPR‐Cas systems in Leptospira reveals potential application of CRISPR in genotyping of Leptospira interrogans

Gao

Che

et al. 2019

APMIS

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“…Thus, at least in some CRISPR-Cas systems, Cas2 might play a secondary role as an RNase, possibly a toxin (101). Indeed, non-sequence-specific nuclease activity of several Cas2 proteins against both DNA and RNA, but typically with a preference for RNA substrates, has been demonstrated although catalytically active Cas2 proteins do not appear to be toxic when overexpressed in E. coli (10, 32, 53, 70, 118). The conservation of the catalytic site of Cas2 implies that the RNase activity of this protein is functional in a subset of CRISPR-Cas systems, and interferase activity remains a distinct possibility.…”

Section: Interactions Between Defense Strategies: Coupling Between Immentioning

confidence: 99%

Evolutionary Genomics of Defense Systems in Archaea and Bacteria

Koonin

Makarova²,

Wolf³

2017

Annu. Rev. Microbiol.

264

220

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Evolution of bacteria and archaea involves an incessant arms race against an enormous diversity of genetic parasites. Accordingly, a substantial fraction of the genes in most bacteria and archaea are dedicated to antiparasite defense. The functions of these defense systems follow several distinct strategies, including innate immunity; adaptive immunity; and dormancy induction, or programmed cell death. Recent comparative genomic studies taking advantage of the expanding database of microbial genomes and metagenomes, combined with direct experiments, resulted in the discovery of several previously unknown defense systems, including innate immunity centered on Argonaute proteins, bacteriophage exclusion, and new types of CRISPR-Cas systems of adaptive immunity. Some general principles of function and evolution of defense systems are starting to crystallize, in particular, extensive gain and loss of defense genes during the evolution of prokaryotes; formation of genomic defense islands; evolutionary connections between mobile genetic elements and defense, whereby genes of mobile elements are repeatedly recruited for defense functions; the partially selfish and addictive behavior of the defense systems; and coupling between immunity and dormancy induction/programmed cell death.

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“…Moreover, the CRISPR/Cas systems also likely harbor a comparable ‘addiction’ component which probably enforces the maintenance of CRISPR/Cas systems in cellular genomes. This component, the Cas2 protein, one of the few core components shared across all CRISPR/Cas systems, targets RNA ( 63 – 65 ).…”

Section: Biological Agents Of Rna Damagementioning

confidence: 99%

RNA damage in biological conflicts and the diversity of responding RNA repair systems

Burroughs

Aravind

2016

Nucleic Acids Res

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RNA is targeted in biological conflicts by enzymatic toxins or effectors. A vast diversity of systems which repair or ‘heal’ this damage has only recently become apparent. Here, we summarize the known effectors, their modes of action, and RNA targets before surveying the diverse systems which counter this damage from a comparative genomics viewpoint. RNA-repair systems show a modular organization with extensive shuffling and displacement of the constituent domains; however, a general ‘syntax’ is strongly maintained whereby systems typically contain: a RNA ligase (either ATP-grasp or RtcB superfamilies), nucleotidyltransferases, enzymes modifying RNA-termini for ligation (phosphatases and kinases) or protection (methylases), and scaffold or cofactor proteins. We highlight poorly-understood or previously-uncharacterized repair systems and components, e.g. potential scaffolding cofactors (Rot/TROVE and SPFH/Band-7 modules) with their respective cognate non-coding RNAs (YRNAs and a novel tRNA-like molecule) and a novel nucleotidyltransferase associating with diverse ligases. These systems have been extensively disseminated by lateral transfer between distant prokaryotic and microbial eukaryotic lineages consistent with intense inter-organismal conflict. Components have also often been ‘institutionalized’ for non-conflict roles, e.g. in RNA-splicing and in RNAi systems (e.g. in kinetoplastids) which combine a distinct family of RNA-acting prim-pol domains with DICER-like proteins.

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Dual nuclease activity of a Cas2 protein in CRISPR–Cas subtype I‐B of Leptospira interrogans

Cited by 27 publications

References 51 publications

Characterization of CRISPR‐Cas systems in Leptospira reveals potential application of CRISPR in genotyping of Leptospira interrogans

Characterization of CRISPR‐Cas systems in Leptospira reveals potential application of CRISPR in genotyping of Leptospira interrogans

Evolutionary Genomics of Defense Systems in Archaea and Bacteria

RNA damage in biological conflicts and the diversity of responding RNA repair systems

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