Inhibition of the B to Z transition in poly(dGdC).poly(dGdC) by covalent attachment of ethidium: equilibrium studies.
The effects of covalent modification of poly(dGdC).poly(dGdC) and poly(dGm5dC).poly(dGm5dC) by ethidium monoazide (a photoreactive analogue of ethidium) on the salt-induced B to Z transition are examined. Earlier studies have shown ethidium monoazide to bind DNA (in the absence of light) in a manner identical to that of the parent ethidium bromide. Photolysis of the ethidium monoazide-DNA complex with visible light results in the covalent attachment of the photoreactive analogue to the DNA. This ability to form a covalent adduct was utilized to probe the effects of an intercalating irreversibly bound adduct on the salt-induced B to Z transition of the poly(dGdC).poly(dGdC) and poly(dGm5dC).poly(dGm5dC) polynucleotides. In the absence of drug, the salt-induced transition from the B to Z structure occurs in a highly cooperative manner. In contrast, this cooperativity is diminished as the concentration of covalently attached drug is increased. The degree of inhibition of the B to Z transition is quantitated as a function of the concentration of covalently attached drug. At a concentration of one drug bound per four base pairs for poly(dGdC).poly(dGdC) and seven base pairs for poly(dGm5dC).poly(dGm5dC), total inhibition of this transition is achieved. Lower concentrations of bound drug were effective in the partial inhibition of this transition. The effects of the covalently bound intercalator on the energetics of the B to Z transition were determined and demonstrated that the adduct is effective in locking the alternating copolymer in a right-handed conformation under high salt conditions.
Gilbert
P L
PL
Department of Chemistry, University of Mississippi, University 38677.
Graves
D E
DE
Chaires
J B
JB
D002151:Calorimetry; D004996:Ethidium / Q000737:chemistry*; D007700:Kinetics; D008433:Mathematics; D009690:Nucleic Acid Conformation; D009994:Osmolar Concentration; D011089:Polydeoxyribonucleotides / Q000737:chemistry*; D012965:Sodium Chloride
D011089:Polydeoxyribonucleotides; C019530:poly(dC-dG); D012965:Sodium Chloride; C029988:poly(dG-m(5)dC).poly(dG-m(5)dC); D004996:Ethidium