Sequence-dependent conformational differences of small RNAs revealed by native gel electrophoresis.
In this study, we use native polyacrylamide gel electrophoresis and one-dimensional NMR spectroscopy to analyze small RNA hairpins containing a UUCG tetraloop. The aggregation state of one RNA 16-mer (5'-CGGCUUCGGUCGACCA-3') in the presence of Mg(2+) was confirmed by laser light scattering. Although it is widely known in the RNA field that some RNAs tend to aggregate, especially when present at high concentrations, the sequence elements responsible for this effect are rarely identified. In this work, we show that Mg(2+)-induced aggregation of the 16-mer RNA hairpin is sensitive to the presence of the 3'-terminal base and a specific 2'-hydroxyl group. Our study highlights the fact that even small changes in a particular RNA sequence can increase its tendency to undergo Mg(2+)-dependent aggregation in an unpredictable manner. Our analysis also shows that native gel electrophoresis is a sensitive probe of RNA conformation with the capability to detect differences apparently caused by subtle base stacking effects at the ends of helices.
Beuning
P J
PJ
Department of Chemistry, University of Minnesota, Minneapolis, Minnesota 55455, USA.
Tessmer
M R
MR
Baumann
C G
CG
Kallick
D A
DA
Musier-Forsyth
K
K
D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D007834:Lasers; D009682:Magnetic Resonance Spectroscopy; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry*; D012542:Scattering, Radiation
D012313:RNA