Effect of DNA-binding drugs on early growth response factor-1 and TATA box-binding protein complex formation with the herpes simplex virus latency promoter.
Adjacent binding sites for early growth response factor-1 (EGR1) and TATA box-binding protein (TBP) were identified on the herpes simplex virus latency promoter in previous work. The binding of EGR1 to the GC-rich region prevented TBP binding to the AT-rich region. With the simultaneous addition of both EGR1 and TBP, the intercalator nogalamycin prevented EGR1 complex formation, resulting in a dose-dependent increase of the TBP.DNA complex. The minor groove binder chromomycin A3 inhibited EGR1 complex formation but resulted in a smaller increase of the TBP complex. In contrast, an alkylating intercalator hedamycin strongly inhibited binding of both proteins. The ability of these GC-binding drugs to prevent EGR1.DNA complex formation was in the following order: hedamycin > nogalamycin > chromomycin A3, and the specificity was nogalamycin > chromomycin A3 > hedamycin. With transcription factor IIA (TFIIA) in the assay, TBP was able to bind the promoter whereas formation of the EGR1.DNA complex was reduced. An AT minor groove-binding drug, distamycin A, disrupted the TBP.TFIIA.DNA complex and restored the EGR1.DNA complex. We conclude that the binding motif and sequence preference of DNA-interactive drugs are manifested in their ability to inhibit the transcription factor-DNA complexes.
Chiang
S Y
SY
Experimental Therapeutics Department, Roswell Park Cancer Institute, Buffalo, New York 14263, USA.
Welch
J J
JJ
Rauscher
F J
FJ
Beerman
T A
TA
D000477:Alkylating Agents / Q000494:pharmacology; D000880:Anthraquinones / Q000494:pharmacology; D001482:Base Composition; D014128:Chromomycin A3 / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000378:metabolism*; D003852:Deoxyribonucleoproteins / Q000737:chemistry; D004214:Distamycins / Q000494:pharmacology; D051766:Early Growth Response Protein 1; D015967:Gene Expression Regulation, Viral; D006801:Humans; D017874:Immediate-Early Proteins; D007364:Intercalating Agents / Q000494:pharmacology*; D046911:Macromolecular Substances; D009621:Nogalamycin / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins; D018139:Simplexvirus / Q000235:genetics*; D035181:TATA-Box Binding Protein; D035561:Transcription Factor TFIIA; D014157:Transcription Factors / Q000378:metabolism*; D017735:Virus Latency
D000477:Alkylating Agents; D000880:Anthraquinones; D004268:DNA-Binding Proteins; D003852:Deoxyribonucleoproteins; D004214:Distamycins; C497325:EGR1 protein, human; D051766:Early Growth Response Protein 1; D017874:Immediate-Early Proteins; D007364:Intercalating Agents; D046911:Macromolecular Substances; D011993:Recombinant Fusion Proteins; D035181:TATA-Box Binding Protein; D035561:Transcription Factor TFIIA; D014157:Transcription Factors; C001328:hedamycin; C030000:stallimycin; D014128:Chromomycin A3; D009621:Nogalamycin