PMID:9792804

Citation	Ahumada, A and Tse-Dinh, YC (1998) The Zn(II) binding motifs of E. coli DNA topoisomerase I is part of a high-affinity DNA binding domain. Biochem. Biophys. Res. Commun. 251:509-14
Abstract	Escherichia coli DNA topoisomerase I binds three Zn(II) with three tetracysteine motifs. Three subclones containing these tetracysteine motifs were expressed and purified. Subclone ZD1 contained the minimal tetracysteine motifs sequence. A larger subclone ZD2 corresponded to a region bordered by two protease sensitive sites. Subclone ZD3 also included the 14-kDa C-terminal domain that has been shown to bind DNA. Subclones ZD1 and ZD2 were found to bind one and two Zn(II), respectively, and neither had detectable DNA binding activity. ZD3 could bind three Zn(II) and had higher DNA binding affinity than the 14-kDa C-terminal domain. The complex formed between ZD3 and a single-stranded 31mer could be detected by the gel shift assay while the complex formed by the 14-kDa C-terminal domain was not stable under gel electrophoresis conditions. The three Zn(II) binding motifs appeared to be part of a high-affinity DNA binding domain.
Links	PubMed Online version:10.1006/bbrc.1998.9500
Keywords	Amino Acid Sequence; Base Sequence; Binding Sites; Catalytic Domain; Cations, Divalent/metabolism; Cloning, Molecular; Cysteine; DNA/metabolism; DNA Primers; DNA Topoisomerases, Type I/chemistry; DNA Topoisomerases, Type I/metabolism; Escherichia coli/enzymology; Kinetics; Polymerase Chain Reaction; Recombinant Proteins/chemistry; Recombinant Proteins/metabolism; Zinc/metabolism
edit table

Significance

You can help EcoliWiki by summarizing why this paper is useful

Useful Materials and Methods

You can help Ecoliwiki by describing the useful materials (strains, plasmids, antibodies, etc) described in this paper.

Annotations

<annotationlinks/>

EcoliWiki Links

Add links to pages that link here (e.g. gene, product, method pages)

References

See Help:References for how to manage references in EcoliWiki.