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PMID: 3011427 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Variations of intramolecular ligation rates allow the detection of protein-induced bends in DNA.

The EMBO journal ·Vol. 5 ·No. 4 ·1986-04-00 ·Pages 799-803

Kotlarz D, Fritsch A, Buc H

Abstract

A method requiring minute amounts of DNA and protein is proposed for the detection of DNA bending in solution. Local bending, induced by the binding of a protein at its stereospecific site, must increase the probability of circularization of short DNA fragments and hence the rate of formation of the corresponding minicircles. This effect should be highly sensitive to the location of the protein-binding site on the DNA fragment. Simple controls allow other possible interpretations to be ruled out. The deformation due to the cyclic AMP receptor protein when it interacts with its stereospecific site at the lactose control region is studied by this method. It is shown in this case that untwisting is negligible and bending significant. Further measurements are required to assess the actual value of the radius of curvature of the DNA double helix in this complex.

MeSH Terms
Base Sequence DNA/metabolism DNA Ligases/metabolism DNA Restriction Enzymes Kinetics Nucleic Acid Conformation Plasmids Promoter Regions, Genetic Protein Binding Receptors, Cyclic AMP/metabolism Structure-Activity Relationship T-Phages/enzymology
Chemicals
Receptors, Cyclic AMP DNA DNA Restriction Enzymes DNA Ligases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kotlarz D
Fritsch A
Buc H
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24 references, click to expand
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1986-04-00
Pages
799-803
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1166861
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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