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XB-ART-26058
Nucleic Acids Res 1990 Feb 25;184:1021-9.
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Reaction parameters of TFIIIA-induced supercoiling catalyzed by a Xenopus laevis cell-free extract.

Sekiguchi JA , Kmiec EB .


Abstract
In addition to its fundamental role of nucleating the formation of stable transcription complexes, the Xenopus laevis 5S RNA specific transcription factor, TFIIIA, promotes a variety of DNA-associated metabolic reactions. We report that TFIIIA can induce a DNA supercoiling catalyzed by the Xenopus laevis S-150 cell-free extract on plasmids containing a single copy of the Xenopus 5S RNA gene (somatic-type). Stimulated supercoiling occurs in the presence of high concentrations of ATP (4 mM) and at a factor to DNA ratio of 1 through a mechanism most likely involving type I topoisomerase. The highest level of stimulated supercoiling occurs when TFIIIA is incubated with DNA prior to the addition of the S-150 extract. Taken together, the experiments outlined in this report establish a reliable and seminal system in which TFIIIA-induced DNA supercoiling can be observed reproducibly.

PubMed ID: 2315014
PMC ID: PMC330359
Article link: Nucleic Acids Res


Species referenced: Xenopus laevis
Genes referenced: gtf3a

References [+] :
Almouzni, Assembly of spaced chromatin promoted by DNA synthesis in extracts from Xenopus eggs. 1988, Pubmed, Xenbase