Cas3-stimulated runaway replication of modified ColE1 plasmids in Escherichia coli is temperature dependent.

08:00 EDT 16th May 2019 | BioPortfolio

Summary of "Cas3-stimulated runaway replication of modified ColE1 plasmids in Escherichia coli is temperature dependent."

The CRISPR-Cas system constitutes an adaptive immunity system of prokaryotes against mobile genetic elements using a crRNA-mediated interference mechanism. In Type I CRISPR-Cas systems, crRNA guided by a Cascade complex recognises the matching target DNA and promotes an R-loop formation, RNA-DNA hybrid. The helicase-nuclease Cas3 protein is then recruited to the Cascade/R-loop complex where it nicks and degrades DNA. The Cas3 activity in CRISPR-Cas immunity is reduced in Δhns cells at 37°C for unknown reasons. Cas3 can also influence regulation of plasmid replication and promote uncontrolled ('runaway') replication of ColE1 plasmids independently of other CRISPR-Cas components, requiring only its helicase activity. In this work we wanted to test whether Cas3-stimulated uncontrolled plasmid replication is affected by the temperature in Δhns and/or ΔhtpG mutants. We found that Cas3-stimulated uncontrolled plasmid replication occurs only at 37°C, irrespective of the genotype of the analysed mutants, and dependent on Cas3 helicase function. We also found that plasmid replication was strongly reduced by the hns mutation at 30°C and that Cas3 could interfere with T4 phage replication at both incubation temperatures.


Journal Details

This article was published in the following journal.

Name: FEMS microbiology letters
ISSN: 1574-6968


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