Binding of distamycin and chromomycin to human immunodeficiency type 1 virus DNA: a non-radioactive automated footprinting study.
Sequence-selectivity of DNA-binding drugs was recently reported in a number of studies employing footprinting and gel retardation approaches. In this paper we studied sequence-selectivity of the binding of chromomycin and distamycin to DNA by performing DNase I footprinting and analysis of the cleaved fragments by the Pharmacia ALF DNA Sequencing System. As a model system we employed the long terminal repeat of the human immunodeficiency type 1 virus. The main conclusion of our experiments is that automated analysis of DNase I footprinting is a fast and reliable technique to study drugs-DNA interactions. The results obtained suggest that distamycin and chromomycin differentially interact with the long terminal repeat of the human immunodeficiency type 1 virus; this differential binding depends upon the DNA sequences recognized. The data presented are consistent with a preferential binding of distamycin to DNA sequences of the binding sites of nuclear factor kappa B and transcription factor IID. By contrast, distamycin exhibits only weak binding to DNA sequences recognized by the promoter-specific transcription factor Sp1. Unlike distamycin, chromomycin preferentially interacts with the binding sites of the promoter-specific transcription factor Sp1.
Feriotto
G
G
Biotechnology Centre, Ferrara University, Italy.
Mischiati
C
C
Bianchi
N
N
Passadore
M
M
Gambari
R
R
D000903:Antibiotics, Antineoplastic / Q000378:metabolism*; D000998:Antiviral Agents / Q000378:metabolism*; D001331:Automation; D001483:Base Sequence; D002865:Chromomycins / Q000378:metabolism*; D018983:DNA Footprinting; D004279:DNA, Viral / Q000378:metabolism*; D003851:Deoxyribonucleases / Q000378:metabolism; D004214:Distamycins / Q000378:metabolism*; D019793:Fluorescein; D005452:Fluoresceins; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
D000903:Antibiotics, Antineoplastic; D000998:Antiviral Agents; D002865:Chromomycins; D004279:DNA, Viral; D004214:Distamycins; D005452:Fluoresceins; C030000:stallimycin; D003851:Deoxyribonucleases; D019793:Fluorescein