Crystallization and Characterization of Pumilio: A Novel RNA Binding Protein |
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Authors: | Thomas A. Edwards Jose Trincao Carlos R. Escalante Robin P. Wharton Aneel K. Aggarwal |
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Affiliation: | a Structural Biology Program, Department of Physiology and Biophysics, Mount Sinai School of Medicine, 1425 Madison Avenue, New York, New York, 10029;b Department of Genetics, Howard Hughes Medical Institute, Duke University Medical Center, Durham, North Carolina, 27710 |
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Abstract: | Axis determination in early Drosophila embryos is controlled, in part, by regulation of translation of mRNAs transcribed in maternal cells during oogenesis. The Pumilio protein is essential in posterior determination, binding to hunchback mRNA in complex with Nanos to suppress hunchback translation. In order to understand the structural basis of RNA binding, Nanos recruitment, and translational control, we have crystallized a domain of the Drosophila Pumilio protein that binds RNA. The crystals belong to the space group P63 with unit cell dimensions of a = b = 94.5 Å, c = 228.9 Å, α = β = 90°, γ = 120° and diffract to 2.6 Å with synchrotron radiation. We show that the purified protein actively binds RNA and is likely to have a novel RNA binding fold due to a very high content of α-helical secondary structure. |
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Keywords: | Pumilio RNA binding protein Nanos crystallization |
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