Determination of the structure of the RNA complex of a double-stranded RNA-binding domain from Drosophila Staufen protein.
We have determined using NMR the structure of the complex between the third double-stranded RNA-binding domain (dsRBD3) of Drosophila Staufen protein and a RNA stem-loop with optimal binding properties in vitro. This work was designed to understand how dsRBD proteins bind RNA and to investigate the role of Staufen dsRBDs in the localization of maternal RNAs during early embryonic development. The structure determination was challenging, because of weak, nonsequence specific binding and residual conformational flexibility at the RNA-protein interface. In order to overcome the problems originated by the weak interaction, we used both new and more traditional approaches to obtain distance and orientation information for the protein and RNA components of the complex. The resulting structure allowed the verification of aspects of RNA recognition by dsRBDs matching the information obtained by a related crystallographic study. We were also able to generate new observations that are likely to be relevant to dsRBD-RNA binding and to the physiological role of Staufen protein.
Ramos
A
A
MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK.
Bayer
P
P
Varani
G
G
D000818:Animals; D001665:Binding Sites; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000737:chemistry; D007301:Insect Hormones / Q000737:chemistry / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D017434:Protein Structure, Tertiary; D012313:RNA / Q000737:chemistry* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism
D029721:Drosophila Proteins; D007301:Insect Hormones; D016601:RNA-Binding Proteins; C069927:stau protein, Drosophila; D012313:RNA