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PMID: 1851553 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Eukaryotic topoisomerase I-DNA interaction is stabilized by helix curvature.

Nucleic acids research ·Vol. 19 ·No. 6 ·1991-03-25 ·Pages 1235-41

Krogh S, Mortensen UH, Westergaard O, Bonven BJ

Abstract

The influence of DNA structure on topoisomerase I-DNA interaction has been investigated using a high affinity binding site and mutant derivatives thereof. Parallel determinations of complex formation and helix structure in the absence of superhelical stress suggest that the interaction is intensified by stable helix curvature. Previous work showed that a topoisomerase I binding site consists of two functionally distinct subdomains. A region located 5' to the topoisomerase I cleavage site is essential for binding. The region 3' to the cleavage site is covered by the enzyme, but not essential. We report here that the helix conformation of the latter region is an important modulator of complex formation. Thus, complex formation is markedly stimulated, when an intrinsically bent DNA segment is installed in this region. A unique pattern of phosphate ethylation interferences in the 3'-part of the binding site indicates that sensing of curvature involves backbone contacts. Since dynamic curvature in supercoiled DNA may substitute for stable curvature, our findings suggest that topoisomerase I is able to probe DNA topology by assessment of writhe, rather than twist.

MeSH Terms
Base Sequence Binding Sites DNA/metabolism DNA Topoisomerases, Type I/genetics,metabolism Electrophoresis, Polyacrylamide Gel Eukaryotic Cells/enzymology Methylation Molecular Sequence Data Nucleic Acid Conformation Plasmids
Chemicals
DNA DNA Topoisomerases, Type I
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Krogh S
Department of Molecular Biology and Plant Physiology, University of Aarhus, Denmark.
Mortensen U H
Westergaard O
Bonven B J
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/19.6.1235
Published
1991-03-25
Pages
1235-41
Language
English
Region
England
NLM ID
0411011
PMCID
PMC333848
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/19.6.1235 PMC Full Text View journal details
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