DNase I footprinting of small molecule binding sites on DNA.
Nuclease footprinting techniques were initially developed to investigate protein-deoxyribonucleic acid (DNA) interactions but these tools of molecular biology have also become instrumental for probing sequence-selective binding of small molecules to DNA. Here, the method is described and technical details are given for performing deoxyribonuclease (DNase) I footprinting with DNA-binding drugs. An example is presented where DNase I is used (as well as DNase II and micrococcal nuclease) to probe the patterns of sequence-selective recognition of DNA by the anticancer antibiotic actinomycin D. DNase I is a convenient endonuclease for detecting and locating the position of actinomycin-binding sites within GC-rich sequences.
Bailly
Christian
C
INSERM U-524, Institut de Recherches sur le Cancer de Lille, France.
Kluza
Jérôme
J
Martin
Christopher
C
Ellis
Thomas
T
Waring
Michael J
MJ
D001345:Autoradiography; D001483:Base Sequence; D001665:Binding Sites; D004247:DNA / Q000378:metabolism*; D018983:DNA Footprinting / Q000379:methods*; D003609:Dactinomycin / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data
D003609:Dactinomycin; D004247:DNA