| Autor: |
Völker, J., Makube, N., Plum, G.E., Klump, H.H., Breslauer, K.J. |
| Předmět: |
|
| Zdroj: |
Proceedings of the National Academy of Sciences of the United States of America; 11/12/2002, Vol. 99 Issue 23, p14700, 6p, 2 Diagrams, 3 Graphs |
| Abstrakt: |
We have embedded the hexameric triplet repeats (CAG)[sub 6] and (CTG)[sub 6] between two (GC)[sub 3] domains to produce two 30-mer hairpins with the sequences d[(GC)[sub 3](CAG)[sub 6](GC)[sub 3]] and d[(GC)[sub 3](CTG)[sub 6](GC)[sub 3]]. This construct reduces the conformational space available to these repetitive DNA sequences. We find that the (CAG)[sub 6] and (CTG)[sub 6] repeats form stable, ordered, single-stranded structures. These structures are stabilized at 62°C by an average enthalpy per base of 1.38 kcal·mol[sup -1] for the CAG triplet and 2.87 kcal·mol[sup -1] for the CTG triplet, while being entropically destabilized by 3.50 cal·K[sup -1]·mol[sup -1] for the CAG triplet and 7.6 cal·K[sup -1]·mol[sup -1] for the CTG triplet. Remarkably, these values correspond, respectively, to ⅓ (for CAG) and 2/3 (for CTG) of the enthalpy and entropy per base values associated with Watson-Crick base pairs. We show that the presence of the loop structure kinetically inhibits duplex formation from the two complementary 30-mer hairpins, even though the duplex is the thermodynamically more stable state. Duplex formation, however, does occur at elevated temperatures. We propose that this thermally induced formation of a more stable duplex results from thermal disruption of the single-stranded order, thereby allowing the complementary domains to associate (perhaps via "kissing hairpins"). Our melting profiles show that, once duplex formation has occurred, the hairpin intermediate state cannot be reformed, consistent with our interpretation of kinetically trapped hairpin structures. The duplex formed by the two complementary oligonucleotides does not have any unusual optical or thermodynamic properties. By contrast, the very stable structures formed by the individual single-stranded triplet repeat sequences are thermally and thermodynamically unusual. We discuss this stable, triplet repeat, single-stranded structure... [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
|
