Studies on DNA Topoisomerase Activity During in Vitro Chromatin Assembly

Overview

Journal Mol Cell Biochem

Publisher Springer

Specialty Biochemistry

Date 1988 Oct 1

PMID 2849054

Citations 2

Authors

J A Sekiguchi

E B Kmiec

Affiliations

Soon will be listed here.

Abstract

The in vitro assembly of chromatin, promoted by the Xenopus cell-free extract (S-150), can be inhibited by oxolinic acid and to a lesser extent by nalidixic acid. Both of these antibiotics have been shown to block the activity of the specialized type II Topoisomerase, bacterial DNA Gyrase. Oxolinic acid induces a DNA cleavage by Micrococcal Nuclease at specific sequences in the multiple cloning vector pGEM-4. Nalidixic acid does not inhibit DNA supercoiling, but does diminish the extent of chromatin formation achieved by the S-150 on circular DNA templates. The Topoisomerase I inhibitor, berenil, does not inhibit extensive chromatin assembly, although it does diminish the level of supercoiling. Taken together, these results suggest that both topoisomerases play a role in the assembly process. Topoisomerase I may catalyze both the introduction of unconstrained supercoils into relaxed DNA and the formation of monosomes, while Topoisomerase II may promote extended chromatin assembly.

Citing Articles

Assembly of nucleosomal DNA in a cell-free extract from wild-type and top1- strains of Ustilago maydis.

Dutta S, Gerhold D, Kmiec E Mol Gen Genet. 1995; 248(6):675-85.

PMID: 7476870 DOI: 10.1007/BF02191707.

In vitro analysis of a type I DNA topoisomerase activity from cultured tobacco cells.

Cole A, Heath-Pagliuso S, BAICH A, Kmiec E Plant Mol Biol. 1992; 19(2):265-76.

PMID: 1320423 DOI: 10.1007/BF00027348.

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