Differential effects of cyclopolyamines on the stability and conformation of triplex DNA.
Linear polyamines are excellent promoters of triplex DNA formation. The effects of structural rigidization of polyamines on triplex DNA stability are not known at present. We wished to develop a series of polyamine analogs as secondary ligands for triplex DNA stabilization for antigene applications. To accomplish this goal, we synthesized cyclopolyamines by interconnecting the two amino or imino groups of linear polyamines with a --(CH2)n-bridge (n=3,4,5). Melting temperature (Tm) data showed that [4,3]-spermine and [4,4]-spermine stabilized poly(dA) x 2poly(dT) triplex at >25 microM concentrations (Tm = 71 degrees C at 100 microM). The dTm/dlog [polyamine] values for these compounds were 26 and 40, respectively. [4,3]-Spermine and [4,4]-spermine also stabilized triplex DNA formed by a purine-motif triplex-forming oligonucleotide, TG3TG4TG4TG3T with its target duplex, as determined by Tm, circular dichroism (CD) spectroscopy, and electrophoretic mobility shift assay (EMSA). In contrast, [4,4]-putrescine and [4,5]-putrescine as well as [4,5]-spermine had no triplex DNA stabilizing effect. CD spectra also showed triplex DNA aggregation and psi-DNA formation at >100 microM [4,3]-spermine. These data demonstrate that structural rigidization of linear polyamines has a profound effect on their ability to stabilize triplex DNA and provoke conformational transitions.
Antony
T
T
Department of Environmental and Community Medicine, University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, New Brunswick 08903, USA.
Musso
M
M
Hosseini
M W
MW
Brand
G
G
Greenfield
N J
NJ
Thomas
T
T
Van Dyke
M W
MW
Thomas
T J
TJ
D002942:Circular Dichroism; D003500:Cyclization; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D008024:Ligands; D009690:Nucleic Acid Conformation / Q000187:drug effects*; D009693:Nucleic Acid Hybridization / Q000187:drug effects; D011061:Poly A / Q000737:chemistry / Q000378:metabolism; D011071:Poly T / Q000737:chemistry / Q000378:metabolism; D011073:Polyamines / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D011687:Purines / Q000737:chemistry / Q000378:metabolism; D011700:Putrescine / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D013095:Spermidine / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D013096:Spermine / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D013696:Temperature; D013816:Thermodynamics
D008024:Ligands; D011073:Polyamines; D011687:Purines; C051802:triplex DNA; D011061:Poly A; D011071:Poly T; D013096:Spermine; D004247:DNA; D013095:Spermidine; D011700:Putrescine