Backbone resonance assignments for a homolog of the essential ribosome biogenesis factor Fap7 from P. horikoshii in its nucleotide-free and -bound forms
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The protein “factor activating Pos9 (Skn7)”, Fap7, is an essential protein in yeast and plays an important role in the biogenesis of the small ribosomal subunit. In eukaryotes, the final processing step of the small ribosomal subunit RNA is the endonucleolytic cleavage of 20S pre-rRNA at cleavage site D yielding mature 18S rRNA. Depletion of Fap7 in yeast leads to a dramatic accumulation of 20S pre-rRNA and a concomitant decrease in 18S rRNA in the cytoplasm. In addition, these cells contain higher levels of 60S, but decreased numbers of 40S ribosomal subunits. Fap7 contains a P-loop like motif placing it in a class with NTPases and kinases and a role for it as an adenylate kinase has been suggested. Up to now both the structure of Fap7 and its detailed function during ribosome biogenesis remain elusive. Here, we present the backbone NMR assignments of a Fap7 homolog from the thermophilic archaeon Pyrococcus horikoshii in its nucleotide free form and bound to the adenylate kinase inhibitor AP5A.
KeywordsNMR-assignments Triple resonance experiments Fap7 Ribosome biogenesis Adenylate kinase
We are grateful to Dr. Elke Duchardt-Ferner for helpful discussions on pulse sequences. This project was supported by the Deutsche Forschungsgemeinschaft (DFG) through the SFB 902 “Molecular mechanisms of RNA-based regulation”, the Center of Excellence Frankfurt (CEF) “Macromolecular complexes”, the Center for Biomolecular Magnetic Resonance Frankfurt (BMRZ) and an Aventis Foundation Professorship (to J.W.).
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