A Study of Triplex Formation of 5′-d-T-(C-T-)₂C-(T-)₄C-(T-C-)₂T with 5′-d-A-(G-A-)₂G A Hairpin Triplex with Three T · A · T and Three C⁺· G · C Bases Triads

Abstract

The UV mixing titration, gel electrophoresis, and CD measurements indicate that oligomers with a basic sequence of 5'-d-T-(C-T-)2C-(T-)4C-(T-C-)2T form a hairpin type triplex with the target 5'-d-A-(G-A-)2G. The stability, measured UV melting temperatures, were studied in aqueous solution as functions of mC (5-methylcytidine) replacement of C, pH (4 to 7), and salt concentrations (up to 1 M). The order of stability is 5'-d-A-(G-A-)2G + 5'-d-T-(C-T-)2C-(T-)4C-(T-C-)2T < 5'-d-A-(G-A-)2G + 5'-d-T-(mC-T-)2mC-(T-)4C-(T-C-)2T approximately 5'-d-A-(G-A-)2G + 5'-d-T-(C-T-)2C-(T-)4 mC-(T-mC-)2T < 5'-d-A-(G-A-)2G + 5'-d-T-(mC-T-)2mC-(T-)4mC-(T-mC-)2T at pH 4. These results indicate that (a) a stable triplex is formed with three T.A.T and three C+.G.C base triads and (b) mC is more effective than C to stablize the triplex formation in acidic condition. Thus, this provides a simple system for further studies of triplex.