Specific inhibition of the transcription factor Ci by a cobalt(III) Schiff base-DNA conjugate.
We describe the use of Co(III) Schiff base-DNA conjugates, a versatile class of research tools that target C2H2 transcription factors, to inhibit the Hedgehog (Hh) pathway. In developing mammalian embryos, Hh signaling is critical for the formation and development of many tissues and organs. Inappropriate activation of the Hedgehog (Hh) pathway has been implicated in a variety of cancers including medulloblastomas and basal cell carcinomas. It is well-known that Hh regulates the activity of the Gli family of C2H2 zinc finger transcription factors in mammals. In Drosophila the function of the Gli proteins is performed by a single transcription factor with an identical DNA binding consensus sequence, Cubitus Interruptus (Ci). We have demonstrated previously that conjugation of a specific 17 base-pair oligonucleotide to a Co(III) Schiff base complex results in a targeted inhibitor of the Snail family C2H2 zinc finger transcription factors. Modification of the oligonucleotide sequence in the Co(III) Schiff base-DNA conjugate to that of Ci's consensus sequence (Co(III)-Ci) generates an equally selective inhibitor of Ci. Co(III)-Ci irreversibly binds the Ci zinc finger domain and prevents it from binding DNA in vitro. In a Ci responsive tissue culture reporter gene assay, Co(III)-Ci reduces the transcriptional activity of Ci in a concentration dependent manner. In addition, injection of wild-type Drosophila embryos with Co(III)-Ci phenocopies a Ci loss of function phenotype, demonstrating effectiveness in vivo. This study provides evidence that Co(III) Schiff base-DNA conjugates are a versatile class of specific and potent tools for studying zinc finger domain proteins and have potential applications as customizable anticancer therapeutics.
Department of Molecular Biosciences, Northwestern University, Evanston, Illinois 60208, USA.
Ryan R
RR
Hurtado
Allison S
AS
Harney
Marie C
MC
Heffern
Robert J
RJ
Holbrook
Robert A
RA
Holmgren
Thomas J
TJ
Meade
D000818:Animals; D002460:Cell Line; D003035:Cobalt / Q000737:chemistry / Q000494:pharmacology*; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors*; D004330:Drosophila; D029721:Drosophila Proteins / Q000037:antagonists & inhibitors*; D050437:Genes, Developmental; D053823:Hedgehog Proteins / Q000037:antagonists & inhibitors*; D006801:Humans; D051379:Mice; D009369:Neoplasms / Q000188:drug therapy; D009841:Oligonucleotides / Q000737:chemistry / Q000494:pharmacology*; D012545:Schiff Bases / Q000737:chemistry / Q000494:pharmacology*; D015398:Signal Transduction; D014157:Transcription Factors / Q000037:antagonists & inhibitors*; D016335:Zinc Fingers
D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D053823:Hedgehog Proteins; D009841:Oligonucleotides; D012545:Schiff Bases; D014157:Transcription Factors; C092937:ci protein, Drosophila; C077245:hh protein, Drosophila; D003035:Cobalt