Structural Changes and Enhancements in DNase I Footprinting Experiments

Jerry Goodisman, James C. Dabrowiak

Research output: Contribution to journalArticle

37 Scopus citations

Abstract

In footprinting experiments, an increase in DNA cleavage with addition of ligand to a system may be due to a ligand-induced structural change. Ligand binding also enhances cleavage by displacing the cleavage agent from ligand-binding sites, thus increasing its concentration elsewhere. The theory and characteristics of this mass-action enhancement are given, and it is shown how it may be recognized. Results of DNase I footprinting of small oligomers, with actinomycin D as ligand, are analyzed to reveal which enhancements are due to mass action, and which can reasonably be ascribed to structural changes. Patterns in the footprinting plots from our experiments on actinomycin D binding to a 139-base-pair DNA fragment (with DNase I as a probe) are studied in the same way. The likely origins of these patterns are discussed, as are enhancements occurring with other probes commonly used in footprinting experiments.

Original languageEnglish (US)
Pages (from-to)1058-1064
Number of pages7
JournalBiochemistry
Volume31
Issue number4
DOIs
StatePublished - Feb 1 1992

ASJC Scopus subject areas

  • Biochemistry

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