title: Nuclear Localization and Gene Expression Modulation by a Fluorescent Sequence-Selective p-Anisyl-benzimidazolecarboxamido Imidazole-Pyrrole Polyamide.
abstract: Synthetic pyrrole (P)-imidazole (I) containing polyamides can target predetermined DNA sequences and modulate gene expression by interfering with transcription factor binding. We have previously shown that rationally designed polyamides targeting the inverted CCAAT box 2 (ICB2) of the topoisomerase IIα (topo IIα) promoter can inhibit binding of transcription factor NF-Y, re-inducing expression of the enzyme in confluent cells. Here, the A/T recognizing fluorophore, p-anisylbenzimidazolecarboxamido (Hx) was incorporated into the hybrid polyamide HxIP, which fluoresces upon binding to DNA, providing an intrinsic probe to monitor cellular uptake. HxIP targets the 5'-TACGAT-3' sequence of the 5' flank of ICB2 with high affinity and sequence specificity, eliciting an ICB2-selective inhibition/displacement of NF-Y. HxIP is readily taken up by NIH3T3 and A549 cells, and detected in the nucleus within minutes. Exposure to the polyamide at confluence resulted in a dose-dependent upregulation of topo IIα expression and enhanced formation of etoposide-induced DNA strand breaks.
authors.affiliation: Cancer Research UK Drug-DNA Interactions Research Group, UCL Cancer Institute, Paul O'Gorman Building, 72 Huntley Street, London WC1E 6BT, UK.
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Kiakos
authors.affiliation: Cancer Research UK Drug-DNA Interactions Research Group, UCL Cancer Institute, Paul O'Gorman Building, 72 Huntley Street, London WC1E 6BT, UK.
authors.forename: Luke
authors.initials: L
authors.lastname: Pett
authors.affiliation: Division of Natural & Applied Sciences and Department of Chemistry, Hope College, 35 East, 12(th) Street, Holland, MI 49423, USA.
authors.forename: Vijay
authors.initials: V
authors.lastname: Satam
authors.affiliation: Division of Natural & Applied Sciences and Department of Chemistry, Hope College, 35 East, 12(th) Street, Holland, MI 49423, USA.
authors.forename: Pravin
authors.initials: P
authors.lastname: Patil
authors.affiliation: Cancer Research UK Drug-DNA Interactions Research Group, UCL Cancer Institute, Paul O'Gorman Building, 72 Huntley Street, London WC1E 6BT, UK.
authors.forename: Daniel
authors.initials: D
authors.lastname: Hochhauser
authors.affiliation: Division of Natural & Applied Sciences and Department of Chemistry, Hope College, 35 East, 12(th) Street, Holland, MI 49423, USA.
authors.forename: Moses
authors.initials: M
authors.lastname: Lee
authors.affiliation: Cancer Research UK Drug-DNA Interactions Research Group, UCL Cancer Institute, Paul O'Gorman Building, 72 Huntley Street, London WC1E 6BT, UK. Electronic address: john.hartley@ucl.ac.uk.
authors.forename: John A
authors.initials: JA
authors.lastname: Hartley
mesh_terms: D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics / Q000378:metabolism; D001562:Benzimidazoles / Q000494:pharmacology; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D015342:DNA Probes / Q000235:genetics / Q000494:pharmacology*; D004250:DNA Topoisomerases, Type II / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005456:Fluorescent Dyes / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D051379:Mice; D041681:NIH 3T3 Cells; D019913:Nuclear Localization Signals / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D009757:Nylons / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011758:Pyrroles / Q000494:pharmacology
chemical_list: D000951:Antigens, Neoplasm; D001562:Benzimidazoles; D015342:DNA Probes; D004268:DNA-Binding Proteins; D005456:Fluorescent Dyes; D007093:Imidazoles; D019913:Nuclear Localization Signals; D009757:Nylons; D011758:Pyrroles; D004250:DNA Topoisomerases, Type II