title: Biochemical characterization of G4 quadruplex telomerase RNA unwinding by the RNA helicase RHAU.
abstract: G4 quadruplexes are stable secondary structures prevalent in DNA and RNA that exhibit diverse regulatory functions. Herein, we describe an in vitro technique using the purified RNA helicase RHAU to unwind a G4 quadruplex identified near the 5' end of the human telomerase RNA (hTR). A synthetic RNA corresponding to the quadruplex forming region of hTR (hTR10-43), as well as a predicted complementary strand (25P1), are combined in a reaction containing the purified helicase and ATP. Reaction products and appropriate controls are resolved by native gel electrophoresis. Gels can be stained using a combination of total RNA and quadruplex-specific dyes to observe the expected quadruplex to duplex conversion. This straightforward method can be extended to study structural changes in other inter- or intramolecular quadruplex containing DNA/RNA molecules with the RHAU helicase or other RNA/DNA remodeling enzymes.
authors.affiliation: Department of Chemistry, University of Manitoba, Room 360, Parker Building, 144 Dysart Road, Winnipeg, MB, Canada, R3T 2N2.
authors.forename: Evan P
authors.initials: EP
authors.lastname: Booy
authors.forename: Ewan K S
authors.initials: EK
authors.lastname: McRae
authors.forename: Sean A
authors.initials: SA
authors.lastname: McKenna
mesh_terms: D053487:DEAD-box RNA Helicases / Q000378:metabolism*; D054856:G-Quadruplexes; D006801:Humans; D012313:RNA / Q000235:genetics*; D019098:Telomerase / Q000235:genetics*
chemical_list: C422952:telomerase RNA; D012313:RNA; D019098:Telomerase; C466988:DHX36 protein, human; D053487:DEAD-box RNA Helicases