期刊
NATURE CHEMICAL BIOLOGY
卷 10, 期 8, 页码 686-U128出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/NCHEMBIO.1561
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资金
- US National Institutes of Health (NIH) [R01-AI081987]
- NIH [T32GM007183]
- US National Institute of General Medical Sciences Medical Scientist National Research Service [5 T32GM07281]
- USA Department of Energy
Spinach is an in vitro-selected RNA aptamer that binds a GFP-like ligand and activates its green fluorescence. Spinach is thus an RNA analog of GFP and has potentially widespread applications for in vivo labeling and imaging. We used antibody-assisted crystallography to determine the structures of Spinach both with and without bound fluorophore at 2.2-angstrom and 2.4-angstrom resolution, respectively. Spinach RNA has an elongated structure containing two helical domains separated by an internal bulge that folds into a G-quadruplex motif of unusual topology. The G-quadruplex motif and adjacent nucleotides comprise a partially preformed binding site for the fluorophore. The fluorophore binds in a planar conformation and makes extensive aromatic stacking and hydrogen bond interactions with the RNA. Our findings provide a foundation for structure-based engineering of new fluorophore-binding RNA aptamers.
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