Sequence specificity of Bacillus subtilis DNA gyrase in vivo

• Published in: Genetica 1991, Vol.85 (1), p.3-12
• Main Authors: BASHKIROV, V. I, ZVINGILA, D. J
• Format: Article
• Language: English
• Subjects: Bacillus subtilis; Bacillus subtilis - enzymology; Bacillus subtilis - genetics; Base Sequence; Binding Sites - genetics; Biological and medical sciences; Consensus Sequence; DNA Topoisomerases, Type II - genetics; DNA Topoisomerases, Type II - metabolism; DNA, Bacterial - drug effects; DNA, Bacterial - genetics; DNA, Bacterial - metabolism; Fundamental and applied biological sciences. Psychology; Genic rearrangement. Recombination. Transposable element; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Oxolinic Acid - pharmacology; Plasmids - genetics; Recombination, Genetic - genetics; Substrate Specificity
• ISSN: 0016-6707; 1573-6857
• DOI: 10.1007/BF00056101

Linearization of pBG0 (a hydrid between Escherichia coli plasmid pBR322 and Staphylococcus aureus plasmid pUB110) was performed by lysis of the oxolinic acid treated Bacillus subtilis protoplasts with sodium dodecyl sulfate. This plasmid DNA linearization was used both for a detailed mapping of DNA gyrase cleavage sites of various strength and for the nucleotide sequence determinations at the points of gyrase-mediated scissions by introducing the XhoI linker DNA. A total of 40 plasmids carrying inserted XhoI linker were sequenced by labeling 3' termini of XhoI sites; 38 of them were found to contain a duplication of four base-pairs of the plasmid sequence flanking the linker, which were characteristic of the oxolinic acid-induced DNA cleavage by E. coli DNA gyrase in vitro and in vivo. The relative strength of these sequenced sites was established by comparing their positions to the sites mapped on the appropriate plasmid genome. This allowed us to propose a consensus sequence of B. subtilis DNA gyrase in vivo cleavage site: [sequence: see text] where N is any nucleotide. The bases in parentheses were preferred secondarily. The involvement of DNA gyrase in illegitimate recombination events in Bacillus subtilis is discussed.