Stability and kinetics of nucleic acid triplexes with chimaeric DNA/RNA third strands

Bernal-Méndez, E.; Leumann, Christian (2002). Stability and kinetics of nucleic acid triplexes with chimaeric DNA/RNA third strands. Biochemistry, 41(41), pp. 12343-12349. American Chemical Society 10.1021/bi025937m

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A series of chimaeric DNA/RNA triplex-forming oligonucleotides (TFOs) with identical base-sequence but varying sequential composition of the sugar residues were prepared. The structural, kinetic and thermodynamic properties of triplex formation with their corresponding double-helical DNA target were investigated by spectroscopic methods. Kinetic and thermodynamic data were obtained from analysis of non-equilibrium UV-melting- and annealing curves in the range of pH 5.1 to 6.7 in a 10 mM citrate/phosphate buffer containing 0.1M NaCl and 1 mM EDTA. It was found that already single substitutions of ribo- for deoxyribonucleotides in the TFOs greatly affect stability and kinetics of triplex formation in a strongly sequence dependent manner. Within the sequence context investigated, triplex stability was found to increase when deoxyribonucleotides were present at the 5'-side and ribonucleotides in the center of the TFO. Especially the substitution of thymidines for uridines in the TFO was found to accelerate both, the association and dissociation process, in a strongly position-dependent way. Differential structural information on triplexes and TFO single-strands was obtained from CD-spectroscopy and gel mobility experiments. Only minor changes were observed in the CD spectra of the triplexes at all pH values investigated, and the electrophoretic mobility was nearly identical in all cases, indicating a high degree of structural similarity. In contrast, the single-stranded TFOs showed high structural variability as determined in the same way. The results are discussed in the context of the design of TFOs for therapeutic or biochemical applications.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Departement of Chemistry and Biochemistry

UniBE Contributor:

Leumann, Christian

Subjects:

500 Science > 570 Life sciences; biology
500 Science > 540 Chemistry

ISSN:

0006-2960

Publisher:

American Chemical Society

Language:

English

Submitter:

Christian Leumann

Date Deposited:

19 May 2015 10:44

Last Modified:

19 May 2015 10:45

Publisher DOI:

10.1021/bi025937m

PubMed ID:

12369823

Uncontrolled Keywords:

ACID analysis Base Sequence CD change chimaeric data DNA kinetic Kinetics methods oligonucleotide Oligonucleotides properties stability sugar thermodynamic thymidine Triplex

BORIS DOI:

10.7892/boris.68583

URI:

https://boris.unibe.ch/id/eprint/68583

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