Identification of RNA-ligand interactions by affinity electrophoresis.
We have developed an affinity electrophoresis method to screen for RNA-ligand interactions. Native polyacrylamide gels were polymerized in the absence and presence of different RNA binding molecules. Binding is indicated by a difference in mobility between the gel with ligand present and the gel with no ligand present. The utility of this method was demonstrated using the known interaction between the Escherichia coli ribosomal A-site RNA and different aminoglycoside ligands. The RNA-aminoglycoside interaction observed is dose dependent, and the affinity mirrors what is observed in solution. In addition, we used this method to gauge the affinity to different aminoglycoside molecules of an RNA molecule derived from the thymidylate synthase mRNA construct that contains a CC mismatch.
Boodram
Sherry N
SN
Department of Chemistry, York University, Toronto, Ontario, Canada M3J 1P3.
Cho
Chul M
CM
Tavares
Tony J
TJ
Johnson
Philip E
PE
D000617:Aminoglycosides / Q000737:chemistry; D020137:Base Pair Mismatch; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D008024:Ligands; D009690:Nucleic Acid Conformation; D012329:RNA, Bacterial / Q000737:chemistry*; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism; D012335:RNA, Ribosomal / Q000737:chemistry; D013940:Thymidylate Synthase / Q000235:genetics / Q000378:metabolism
D000617:Aminoglycosides; D008024:Ligands; D012329:RNA, Bacterial; D012333:RNA, Messenger; D012335:RNA, Ribosomal; D013940:Thymidylate Synthase