Stabilization of a DNA duplex under molecular crowding conditions of PEG.

Abstract

A living cell generally contains macromolecules occupying 20-40% of the total volume. To mimic the crowded cellular condition, we prepared solutions including poly(ethylene glycol) (PEG) as a cosolute and investigated the influence of the cosolute on the DNA duplex stability. In the presence of PEG 200 or PEG 8000, the Tm (melting temperature) of a self-complementary duplex of 5'-dATGCGCAT-3' decreased by 11.8 degrees C in the presence of 20 wt% PEG200 and by 1.5 degrees C in the presence of 20 wt% PEG 8000. The dln K(obs) vs. dln a(w) plots for PEG 200 and PEG 8000 were linear with a negative slope, suggesting the association of water molecules upon the duplex formation. Interestingly, when the NaCl concentration decreased from 1 M to 400 mM, the Tm increased in the presence of PEGs. Our results imply that the nucleic acid stabilities in a living cell may be different from those in in vitro conditions.