Specific inhibition of the transcription factor Ci by a cobalt(III) Schiff base-DNA conjugate

Ryan R. Hurtado, Allison S. Harney, Marie C. Heffern, Robert J. Holbrook, Robert A Holmgren, Thomas J Meade*

*Corresponding author for this work

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)325-333
Number of pages9
JournalMolecular Pharmaceutics
Volume9
Issue number2
DOIs
StatePublished - Feb 6 2012

Fingerprint

Schiff Bases
Cobalt
Hedgehogs
Transcription Factors
DNA
Zinc Fingers
Consensus Sequence
Oligonucleotides
Drosophila
Mammalian Embryo
Medulloblastoma
Basal Cell Carcinoma
Snails
Reporter Genes
Base Pairing
Mammals
Embryonic Structures
Phenotype
Injections
Research

Keywords

  • Cubitus Interruptus
  • Drosophila
  • Hedgehog signaling
  • basal cell carcinoma
  • cobalt chelate/Schiff base
  • development
  • transcription factor
  • zinc fingers

ASJC Scopus subject areas

  • Pharmaceutical Science
  • Molecular Medicine
  • Drug Discovery

Cite this

Hurtado, Ryan R. ; Harney, Allison S. ; Heffern, Marie C. ; Holbrook, Robert J. ; Holmgren, Robert A ; Meade, Thomas J. / Specific inhibition of the transcription factor Ci by a cobalt(III) Schiff base-DNA conjugate. In: Molecular Pharmaceutics. 2012 ; Vol. 9, No. 2. pp. 325-333.
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abstract = "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.",
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Specific inhibition of the transcription factor Ci by a cobalt(III) Schiff base-DNA conjugate. / Hurtado, Ryan R.; Harney, Allison S.; Heffern, Marie C.; Holbrook, Robert J.; Holmgren, Robert A; Meade, Thomas J.

In: Molecular Pharmaceutics, Vol. 9, No. 2, 06.02.2012, p. 325-333.

Research output: Contribution to journalArticle

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AU - Hurtado, Ryan R.

AU - Harney, Allison S.

AU - Heffern, Marie C.

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AU - Meade, Thomas J

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