Binding of Triple Helix Forming Oligonucleotides to Sites in Gene Promoters

Ross H. Durland, Donald J. Kessler, Sandy Gunnell, Michael E. Hogan, Madeleine Duvic, B. M. Pettitt

Research output: Contribution to journalArticlepeer-review

253 Scopus citations

Abstract

A class of triplex-forming oligodeoxyribonucleotides (TFOs) is described that can bind to naturally occurring sites in duplex DNA at physiological pH in the presence of magnesium. The data are consistent with a structure in which the TFO binds in the major groove of double-stranded DNA to form a three-stranded complex that is superficially similar to previously described triplexes. The distinguishing features of this class of triplex are that TFO binding apparently involves the formation of hydrogen-bonded G·GC and T·AT triplets and the TFO is bound antiparallel with respect to the more purine-rich strand of the underlying duplex. Triplex formation is described for targets in the promoter regions of three different genes: the human c-myc and epidermal growth factor receptor genes and the mouse insulin receptor gene. All three sites are relatively GC rich and have a high percentage of purine residues on one strand. DNase I footprinting shows that individual TFOs bind selectively to their target sites at pH 7.4–7.8 in the presence of millimolar concentrations of magnesium. Electrophoretic analysis of triplex formation indicates that specific TFOs bind to their target sites with apparent dissociation constants in the 10−7–10−9 M range. Strand orientation of the bound TFOs was confirmed by attaching eosin or an iron-chelating group to one end of the TFO and monitoring the pattern of damage to the bound duplex DNA. Possible hydrogen-bonding patterns and triplex structures are discussed.

Original languageEnglish (US)
Pages (from-to)9246-9255
Number of pages10
JournalBiochemistry
Volume30
Issue number38
DOIs
StatePublished - Sep 1 1991
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

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