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Int. J. Mol. Sci. 2017, 18(7), 1340; doi:10.3390/ijms18071340

Reverse Gyrase Functions in Genome Integrity Maintenance by Protecting DNA Breaks In Vivo

Archaeal Centre, Department of Biology, University of Copenhagen, Ole Maaløes Vej 5, Copenhagen Biocenter, DK-2200 Copenhagen N, Denmark
Authors to whom correspondence should be addressed.
Received: 27 May 2017 / Revised: 14 June 2017 / Accepted: 20 June 2017 / Published: 22 June 2017
(This article belongs to the Special Issue DNA Injury and Repair Systems)
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Reverse gyrase introduces positive supercoils to circular DNA and is implicated in genome stability maintenance in thermophiles. The extremely thermophilic crenarchaeon Sulfolobus encodes two reverse gyrase proteins, TopR1 (topoisomerase reverse gyrase 1) and TopR2, whose functions in thermophilic life remain to be demonstrated. Here, we investigated the roles of TopR1 in genome stability maintenance in S. islandicus in response to the treatment of methyl methanesulfonate (MMS), a DNA alkylation agent. Lethal MMS treatment induced two successive events: massive chromosomal DNA backbone breakage and subsequent DNA degradation. The former occurred immediately after drug treatment, leading to chromosomal DNA degradation that concurred with TopR1 degradation, followed by chromatin protein degradation and DNA-less cell formation. To gain a further insight into TopR1 function, the expression of the enzyme was reduced in S. islandicus cells using a CRISPR-mediated mRNA interference approach (CRISPRi) in which topR1 mRNAs were targeted for degradation by endogenous III-B CRISPR-Cas systems. We found that the TopR1 level was reduced in the S. islandicus CRISPRi cells and that the cells underwent accelerated genomic DNA degradation during MMS treatment, accompanied by a higher rate of cell death. Taken together, these results indicate that TopR1 probably facilitates genome integrity maintenance by protecting DNA breaks from thermo-degradation in vivo. View Full-Text
Keywords: reverse gyrase; Sulfolobus; MMS; genomic DNA breakage; genomic DNA degradation; CRISPRi approach reverse gyrase; Sulfolobus; MMS; genomic DNA breakage; genomic DNA degradation; CRISPRi approach

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Han, W.; Feng, X.; She, Q. Reverse Gyrase Functions in Genome Integrity Maintenance by Protecting DNA Breaks In Vivo. Int. J. Mol. Sci. 2017, 18, 1340.

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