2014
DOI: 10.1093/nar/gku384
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Increased negative supercoiling of mtDNA in TOP1mt knockout mice and presence of topoisomerases II  and II  in vertebrate mitochondria

Abstract: Topoisomerases are critical for replication, DNA packing and repair, as well as for transcription by allowing changes in DNA topology. Cellular DNA is present both in nuclei and mitochondria, and mitochondrial topoisomerase I (Top1mt) is the only DNA topoisomerase specific for mitochondria in vertebrates. Here, we report in detail the generation of TOP1mt knockout mice, and demonstrate that mitochondrial DNA (mtDNA) displays increased negative supercoiling in TOP1mt knockout cells and murine tissues. This find… Show more

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Cited by 73 publications
(109 citation statements)
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References 51 publications
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“…Each KO mouse had at least one sibling WT as control. Mice were genotyped by PCR using genomic DNA from mice tail tips (5). Three primers were used as follows: TOP1mt-A (5′-GGTGCTA-GACATTGAACTCAG-), TOP1mt-B (5′-CTGCAAATGGCCTCGTTAGC), and TOP1mt-C (5′-GTCCTGGATTCCATCTTAAGC).…”
Section: Methodsmentioning
confidence: 99%
See 1 more Smart Citation
“…Each KO mouse had at least one sibling WT as control. Mice were genotyped by PCR using genomic DNA from mice tail tips (5). Three primers were used as follows: TOP1mt-A (5′-GGTGCTA-GACATTGAACTCAG-), TOP1mt-B (5′-CTGCAAATGGCCTCGTTAGC), and TOP1mt-C (5′-GTCCTGGATTCCATCTTAAGC).…”
Section: Methodsmentioning
confidence: 99%
“…To address the role of mtDNA in liver regeneration, we took advantage of the fact that mice lacking TOP1mt are viable (5,11,17). We suspected that Top1mt might play a role in liver regeneration because we recently uncovered a critical role for TOP1mt in maintaining mtDNA homeostasis and mitochondrial function in adaptive response to doxorubicin-induced cardiotoxicity (17).…”
mentioning
confidence: 99%
“…The protein has a molecular mass of about 72 kDa and can relax supercoiled DNA generated by replication or transcription (106,107). Recently, two nuclear type IIA topoisomerases, TOP2α and TOP2β, which catalyzes transient double-stranded DNA breaks, were found to be present and active in mammalian mitochondria (108). Interestingly, TOP2α forms a complex with mtDNA at both ends of 7S DNA and may thus be involved in regulating the stability of the D-loop structure.…”
Section: The Mtdna Replisomementioning
confidence: 99%
“…Before termination of daughter strand replication, the two mtDNA must segregate to avoid catenation. A recent study of human breast cancer and osteosarcoma cell lines has determined that the type IIA topoisomerase, Top2α is the most prevalent human mitochondrial gyrase critical for decatenation of mtDNA circles during replication and relaxation of positive supercoils introduced during transcription and mtDNA replication [33]. …”
Section: Overview Of Human Mtdna Replicationmentioning
confidence: 99%