期刊
JOURNAL OF BIOLOGICAL CHEMISTRY
卷 288, 期 49, 页码 35014-35027出版社
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
DOI: 10.1074/jbc.M113.512970
关键词
DNA Helicase; RNA-binding Protein; RNA Helicase; RNA-Protein Interaction; X-ray Scattering; DHX36; G-quadruplex; G4R1; RHAU
资金
- Canadian Institutes of Health Research (CIHR)/Manitoba Health Research Council (MHRC) Regional Partnership Program [RPA-118069]
- CIHR postdoctoral fellowship
- MHRC postdoctoral fellowship
- University of Manitoba Faculty of Science undergraduate student research award
- University of Manitoba undergraduate research fellowship
- University of Manitoba graduate fellowship
- Canada Research Chair program
Background: The helicase RHAU requires an N-terminal extension to bind quadruplex structures. Results: This extension adopts an elongated shape and interacts with the guanine tetrad face of quadruplexes. Conclusion: We provide a basis for the understanding of quadruplex binding by the N-terminal domain. Significance: The N-terminal region does not require the 2-OH of the ribose to mediate the protein-quadruplex interaction. Polynucleotides containing consecutive tracts of guanines can adopt an intramolecular G-quadruplex structure where multiple planar tetrads of hydrogen-bound guanines stack on top of each other. Remodeling of G-quadruplexes impacts numerous aspects of nucleotide biology including transcriptional and translational control. RNA helicase associated with AU-rich element (RHAU), a member of the ATP-dependent DEX(H/D) family of RNA helicases, has been established as a major cellular quadruplex resolvase. RHAU contains a core helicase domain responsible for ATP binding/hydrolysis/helicase activity and is flanked on either side by N- and C-terminal extensions. The N-terminal extension is required for quadruplex recognition, and we have previously demonstrated complex formation between this domain and a quadruplex from human telomerase RNA. Here we used an integrated approach that includes small angle x-ray scattering, nuclear magnetic resonance spectroscopy, circular dichroism, and dynamic light scattering methods to demonstrate the recognition of G-quadruplexes by the N-terminal domain of RHAU. Based on our results, we conclude that (i) quadruplex from the human telomerase RNA and its DNA analog both adopt a disc shape in solution, (ii) RHAU(53-105) adopts a defined and extended conformation in solution, and (iii) the N-terminal domain mediates an interaction with a guanine tetrad face of quadruplexes. Together, these data form the foundation for understanding the recognition of quadruplexes by the N-terminal domain of RHAU.
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