Distamycins inhibit the binding of OTF-1 and NFE-1 transfactors to their conserved DNA elements.
We investigated the effects of the antiviral agent distamycin A and of a distamycin derivative (FCE 24517) which possesses antineoplastic activity on the binding of some regulatory proteins to DNA. Both compounds inhibited the binding to DNA of the ubiquitous octamer binding factor OTF 1 and of the erythroid specific GATAAG protein (NFE 1). This was shown using the electrophoretic mobility shift assay on a DNA fragment of human gamma-globin gene promoter (-156 to -201), on the same fragment with a point mutation (T to C mutation) known to have an increased affinity of binding for NFE 1, on a DNA fragment of human histone H2B promoter and on a DNA fragment of mouse alpha 1 globin promoter. The ability of distamycin or of FCE 24517 to inhibit the binding was specific for AT-rich sequences since neither drug inhibited the binding of nuclear protein factors to the sequence CCACACCC of the human beta globin gene. Binding to DNA was investigated by evaluating the drugs' ability to protect selected sequences from DNase I digestion (DNase footprinting). Distamycins binding was highly preferential for DNA sequences containing stretches of AT. These studies indicate that chemicals which have a high degree of DNA sequence-specific binding can selectively inhibit the binding of regulatory proteins to DNA. These effects might be responsible for modification of the transcription of specific genes and might to some extent account for these drugs' antiviral and antineoplastic activities. This approach offers potential for the investigation of new such drugs.
Broggini
M
M
Laboratory of Cancer Chemotherapy, Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy.
Ponti
M
M
Ottolenghi
S
S
D'Incalci
M
M
Mongelli
N
N
Mantovani
R
R
D001483:Base Sequence / Q000187:drug effects; D001667:Binding, Competitive; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004214:Distamycins / Q000378:metabolism / Q000494:pharmacology*; D006657:Histones / Q000378:metabolism; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000235:genetics / Q000378:metabolism; D009841:Oligonucleotides / Q000378:metabolism; D011758:Pyrroles / Q000494:pharmacology*; D014157:Transcription Factors / Q000378:metabolism*
D004268:DNA-Binding Proteins; D004214:Distamycins; D006657:Histones; D009841:Oligonucleotides; D011758:Pyrroles; D014157:Transcription Factors