title: The DEAH-box RNA helicase RHAU binds an intramolecular RNA G-quadruplex in TERC and associates with telomerase holoenzyme.
abstract: Guanine-quadruplexes (G4) consist of non-canonical four-stranded helical arrangements of guanine-rich nucleic acid sequences. The bulky and thermodynamically stable features of G4 structures have been shown in many respects to affect normal nucleic acid metabolism. In vivo conversion of G4 structures to single-stranded nucleic acid requires specialized proteins with G4 destabilizing/unwinding activity. RHAU is a human DEAH-box RNA helicase that exhibits G4-RNA binding and resolving activity. In this study, we employed RIP-chip analysis to identify en masse RNAs associated with RHAU in vivo. Approximately 100 RNAs were found to be associated with RHAU and bioinformatics analysis revealed that the majority contained potential G4-forming sequences. Among the most abundant RNAs selectively enriched with RHAU, we identified the human telomerase RNA template TERC as a true target of RHAU. Remarkably, binding of RHAU to TERC depended on the presence of a stable G4 structure in the 5'-region of TERC, both in vivo and in vitro. RHAU was further found to associate with the telomerase holoenzyme via the 5'-region of TERC. Collectively, these results provide the first evidence that intramolecular G4-RNAs serve as physiologically relevant targets for RHAU. Furthermore, our results suggest the existence of alternatively folded forms of TERC in the fully assembled telomerase holoenyzme.
authors.affiliation: Friedrich Miescher Institute for Biomedical Research, Novartis Research Foundation, Maulbeerstrasse 66, 4058 Basel, Switzerland. simon.lattmann@fmi.ch
authors.forename: Simon
authors.initials: S
authors.lastname: Lattmann
authors.forename: Michael B
authors.initials: MB
authors.lastname: Stadler
authors.forename: James P
authors.initials: JP
authors.lastname: Vaughn
authors.forename: Steven A
authors.initials: SA
authors.lastname: Akman
authors.forename: Yoshikuni
authors.initials: Y
authors.lastname: Nagamine
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D001665:Binding Sites; D002460:Cell Line; D053487:DEAD-box RNA Helicases / Q000378:metabolism*; D054856:G-Quadruplexes; D020035:Holoenzymes / Q000378:metabolism; D006801:Humans; D059372:Nucleotide Motifs; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000737:chemistry* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000378:metabolism; D019098:Telomerase / Q000737:chemistry / Q000378:metabolism*
chemical_list: D020035:Holoenzymes; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; C422952:telomerase RNA; D012313:RNA; D019098:Telomerase; D000251:Adenosine Triphosphatases; C466988:DHX36 protein, human; D053487:DEAD-box RNA Helicases