Hairpin ribozyme-catalyzed ligation in water-alcohol solutions.
The hairpin ribozyme (HPR) is a naturally existing RNA that catalyzes site-specific RNA cleavage and ligation. At 37 degrees C and in the presence of divalent metal ions (M(2+)), the HPR efficiently cleaves RNA substrates in trans. Here, we show that the HPR can catalyze efficient M(2+)-independent ligation in trans in aqueous solutions containing any of several alcohols, including methanol, ethanol, and isopropanol, and millimolar concentrations of monovalent cations. Ligation proceeds most efficiently in 60% isopropanol at 37 degrees C, whereas the reverse (cleavage) reaction is negligible under these conditions. We suggest that dehydration of the RNA is the key factor promoting HPR activity in water- alcohol solutions. Alcohol-induced ribozyme ligation may have practical applications.
Vlassov
Alexander V
AV
SomaGenics, Inc., Santa Cruz, CA 95060, USA. avlassov@somagenics.com
Johnston
Brian H
BH
Kazakov
Sergei A
SA
D000438:Alcohols / Q000737:chemistry; D001483:Base Sequence; D002384:Catalysis; D017931:DNA Primers; D007700:Kinetics; D009690:Nucleic Acid Conformation; D016337:RNA, Catalytic / Q000737:chemistry / Q000378:metabolism*; D012996:Solutions; D013696:Temperature; D014867:Water / Q000737:chemistry*
D000438:Alcohols; D017931:DNA Primers; D016337:RNA, Catalytic; D012996:Solutions; D014867:Water