Polyamines alter sequence-specific DNA-protein interactions.
The polyamines are abundant biogenic cations implicated in many biological processes. Despite a plethora of evidence on polyamine-induced DNA conformational changes, no thorough study of their effects on the activities of sequence-specific DNA binding proteins has been performed. We describe the in vitro effects of polyamines on the activities of purified, representative DNA-binding proteins, and on complex protein mixtures. Polyamines at physiological concentrations enhance the binding of several proteins to DNA (e.g. USF, TFE3, Ig/EBP, NF-IL6, YY1 and ICP-4, a herpes simplex virus gene regulator), but inhibit others (e.g. Oct-1). The degree of enhancement correlates with cationic charge; divalent putrescine is ineffective whereas tetravalent spermine is more potent than trivalent spermidine. Polyamine effects on USF and ICP-4 result from increased rate of complex formation rather than a decreased rate of dissociation. DNAse I footprint analysis indicated that polyamines do not alter DNA-protein contacts. Polyamines also facilitate formation of complexes involving binding of more than one protein on a DNA fragment.
Panagiotidis
C A
CA
Department of Microbiology, College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.
Artandi
S
S
Calame
K
K
Silverstein
S J
SJ
D001483:Base Sequence; D003035:Cobalt / Q000494:pharmacology; D004247:DNA / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D006367:HeLa Cells; D018259:Herpesvirus 1, Human / Q000235:genetics; D006801:Humans; D017874:Immediate-Early Proteins / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D011073:Polyamines / Q000494:pharmacology*; D011485:Protein Binding; D011700:Putrescine / Q000494:pharmacology; D011838:Radiation-Sensitizing Agents / Q000494:pharmacology; D013095:Spermidine / Q000494:pharmacology; D013096:Spermine / Q000494:pharmacology; D013379:Substrate Specificity
D004268:DNA-Binding Proteins; D017874:Immediate-Early Proteins; D009838:Oligodeoxyribonucleotides; D011073:Polyamines; D011838:Radiation-Sensitizing Agents; C048925:herpes simplex virus, type 1 protein ICP4; C032300:hexaamminecobalt(II); D013096:Spermine; D003035:Cobalt; D004247:DNA; D013095:Spermidine; D011700:Putrescine