Design, synthesis, and analysis of conformationally constrained nucleic acids

Area of Science:

• Biochemistry
• Molecular Biology
• Chemical Biology

Background:

• Proteins are extensively studied using cystine cross-links to probe structure, dynamics, thermodynamics, folding, and function.
• A similar approach is needed for nucleic acids (DNA and RNA) to enhance structural and functional studies.

Purpose of the Study:

• To review straightforward and general methods for modifying nucleic acid structures using disulfide cross-links.
• To highlight the utility of disulfide cross-links as tools for studying nucleic acid properties.

Main Methods:

• Discussion of methods to introduce disulfide cross-links into nucleic acid structures.
• Synthesis of various nucleic acid constructs (hairpins, duplexes, triplexes, tRNAs) with disulfide cross-links.

Main Results:

• Disulfide cross-links can be introduced quantitatively under mild air oxidation conditions.
• The cross-linking modification does not perturb nucleic acid secondary or tertiary structure.
• Successfully synthesized disulfide cross-linked nucleic acid structures including hairpins, duplexes, triplexes, and tRNAs.

Conclusions:

• Disulfide cross-linking is a versatile and non-perturbing method for studying nucleic acids.
• This chemistry provides a valuable tool for investigating nucleic acid structure, dynamics, thermodynamics, folding, and function.