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A dual function of the CRISPR–Cas system in bacterial antivirus immunity and DNA repair
Abstract
Summary: Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs) and the associated proteins (Cas) comprise a system of adaptive immunity against viruses and plasmids in prokaryotes. Cas1 is a CRISPR‐associated protein that is common to all CRISPR‐containing prokaryotes but its function remains obscure. Here we show that the purified Cas1 protein of Escherichia coli (YgbT) exhibits nuclease activity against single‐stranded and branched DNAs including Holliday junctions, replication forks and 5′‐flaps. The crystal structure of YgbT and site‐directed mutagenesis have revealed the potential active site. Genome‐wide screens show that YgbT physically and genetically interacts with key components of DNA repair systems, including recB , recC and ruvB . Consistent with these findings, the ygbT deletion strain showed increased sensitivity to DNA damage and impaired chromosomal segregation. Similar phenotypes were observed in strains with deletion of CRISPR clusters, suggesting that the function of YgbT in repair involves interaction with the CRISPRs. These results show that YgbT belongs to a novel, structurally distinct family of nucleases acting on branched DNAs and suggest that, in addition to antiviral immunity, at least some components of the CRISPR–Cas system have a function in DNA repair.
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Both Cas1 and Cas2 have nuclease activity.
“Both proteins have nuclease activity [21,22]”
From CRISPR-Cas: Converting A Bacterial Defence Mechanism into A State-of-the-Art Genetic Manipulation Tool (2019) · cites this paper as [21]
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“coli, we constructed chromosomal yellow fluorescent protein (YFP) fusions (Taniguchi et al., 2010) in which YgbT, RuvA, RuvB and RuvC genes were tagged with the YFP-coding sequence at their C-terminal ends.”
BackgroundMethodSelf-citationInteraction network containing conserved and essential protein complexes in Escherichia coli
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Self-citationReplication fork stalling at natural impediments
Mirkin, E. V., & Mirkin, S. M. (2007) · Microbiology and Molecular Biology Reviews · doi:10.1128/mmbr.00030-06 · Cited by 482
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RNA-guided genetic silencing systems in bacteria and archaea
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CRISPR-Cas Systems in Bacteria and Archaea: Versatile Small RNAs for Adaptive Defense and Regulation
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Metadata
- Authors
- Babu, M., Beloglazova, N., Flick, R., Graham, C., Skarina, T., Nocek, B., Gagarinova, A., Pogoutse, O., Brown, G., Binkowski, A., Phanse, S., Joachimiak, A., Koonin, E. V., Savchenko, A., Emili, A., Greenblatt, J., Edwards, A. M., & Yakunin, A. F.
- Venue
- Molecular Microbiology
- Publisher
- Wiley
- Published
- 2010-12-07
- Volume
- 79
- Issue
- 2
- Pages
- 484-502
- ISSN
- 0950-382X
- 1365-2958
- PMC
- PMC3071548
- Cited by
- 250
- References
- 3
- Language
- en
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