Kinetics and Thermodynamics of DNA, RNA, and Hybrid Duplex Formation

✍ Scribed by Rauzan, Brittany; McMichael, Elizabeth; Cave, Rachel; Sevcik, Lesley R.; Ostrosky, Kara; Whitman, Elisabeth; Stegemann, Rachel; Sinclair, Audra L.; Serra, Martin J.; Deckert, Alice A.

• Book ID: 121331591
• Publisher: American Chemical Society
• Year: 2013
• Tongue: English
• Weight: 541 KB
• Volume: 52
• Category: Article
• ISSN: 0006-2960
• DOI: 10.1021/bi3013005

✦ Synopsis

The rates of duplex formation for two octamers of DNA (5′ d-CACGGCTC/5′ d-GAGCCGTG and 5′ d-CACAGCAC/5′ d-GTGCTGTG), the homologous RNA, and both sets of hybrids in 1 M NaCl buffer have been measured using stopped-flow spectroscopy. In addition, the thermodynamic parameters, ΔH°and ΔS°, have been determined for the same sequences under the same buffer conditions using optical melting techniques. These data reveal a linear free energy relationship between the free energy of activation for denaturation and the change in free energy for formation of the duplexes. This relationship indicates that these duplex formation reactions occur through a common unstructured transition state that is more similar to the single strands in solution than to the ensuing duplex. In addition, these data confirm that the greater stability of RNA duplexes relative to that of homologous DNA and hybrid duplexes is controlled by the denaturation rate and not the duplex formation rate.