Precise mapping of RNA tertiary structure via nanometer distance measurements with double electron-electron resonance spectroscopy.
Divalent metal (Mg(2+)) ion-dependent folding of the hammerhead ribozyme from Schistosoma mansoni was monitored with double electron-electron resonance (DEER) pulsed electron paramagnetic resonance spectroscopy by measuring nanometer-scale distances between paramagnetic spin-labels attached to the RNA. DEER measurements detect global folding of the ribozyme with excellent agreement between predictions from experimental, modeled, and crystallographic measurements. These measurements demonstrate the use of DEER spectroscopy as a tool for structural analysis of complex RNAs.
Kim
Nak-Kyoon
NK
Department of Chemistry, Texas A&M University, College Station, Texas 77842, USA.
Bowman
Michael K
MK
DeRose
Victoria J
VJ
D000818:Animals; D001483:Base Sequence; D018360:Crystallography, X-Ray; D004583:Electrons; D008670:Metals / Q000494:pharmacology; D009690:Nucleic Acid Conformation / Q000187:drug effects; D012313:RNA / Q000737:chemistry* / Q000235:genetics; D016337:RNA, Catalytic / Q000737:chemistry / Q000235:genetics; D012550:Schistosoma mansoni / Q000201:enzymology; D013057:Spectrum Analysis
D008670:Metals; D016337:RNA, Catalytic; C427204:hammerhead ribozyme; D012313:RNA