Triplex formation at the rat neu gene utilizing imidazole and 2'-deoxy-6-thioguanosine base substitutions.
Triplex-forming oligodeoxyribonucleotides (TFOs) can be designed so as to form antiparallel triple helices with duplex DNA by means of GGC and TAT or AAT base triplets, and these have been shown to be useful as sequence-specific DNA binding agents. Using TFOs targeted to the promoter region of the rat neu oncogene, it is shown here that substitution of an imidazole-nucleoside chimera at a single site in a neu specific TFO results in an increase in TFO binding affinity and specificity. This effect is discussed in terms of the stabilizing effect of local imidazole-TA triplet formation. It is also found that site-selective substitution of 2'-deoxy-6-thioguanosine for guanosine (S6-dG) in the TFO results in an increase in triplex formation in the presence of physiological levels of potassium ion. The utility and positioning of S6-dG base substitutions is discussed in the context of an intramolecular tetrad model.
Center for Biotechnology, Baylor College of Medicine, Woodlands, Texas 77381.
J E
JE
Gee
G R
GR
Revankar
T S
TS
Rao
M E
ME
Hogan
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000737:chemistry; D003850:Deoxyribonuclease I; D018734:Genes, erbB-2; D007093:Imidazoles / Q000737:chemistry*; D008274:Magnesium / Q000494:pharmacology; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides / Q000737:chemistry*; D051381:Rats; D013329:Structure-Activity Relationship; D013872:Thionucleosides / Q000737:chemistry*
D007093:Imidazoles; D009838:Oligodeoxyribonucleotides; D013872:Thionucleosides; C010438:beta-2'-deoxythioguanosine; D003850:Deoxyribonuclease I; D003849:Deoxyguanosine; D008274:Magnesium