Biomolecular NMR Assignments

, Volume 7, Issue 2, pp 261–265 | Cite as

Backbone resonance assignments for a homolog of the essential ribosome biogenesis factor Fap7 from P. horikoshii in its nucleotide-free and -bound forms

  • Ute A. Hellmich
  • Jens WöhnertEmail author


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.


NMR-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.).


  1. Gavin AC, Bosche M, Krause R, Grandi P, Marzioch M et al (2002) Functional organization of the yeast proteome by systematic analysis of protein complexes. Nature 415:141–147ADSCrossRefGoogle Scholar
  2. Granneman S, Nandineni MR, Baserga SJ (2005) The putative NTPase Fap7 mediates cytoplasmic 20S pre-rRNA processing through a direct interaction with Rps14. Mol Cell Biol 25:10352–10364CrossRefGoogle Scholar
  3. Hellmich UA, Duchardt-Ferner E, Glaubitz C, Wöhnert J (2012) Backbone NMR resonance assignments of the nucleotide binding domain of the ABC multidrug transporter LmrA from Lactococcus lactis in its ADP-bound state. Biomol NMR Assign 6:69–73CrossRefGoogle Scholar
  4. Jakovljevic J, de Mayolo PA, Miles TD, Nguyen TM, Léger-Silvestre I et al (2004) The carboxy-terminal extension of yeast ribosomal protein S14 is necessary for maturation of 43S preribosomes. Mol Cell 14:331–342CrossRefGoogle Scholar
  5. Juhnke H, Charizanis C, Latifi F, Krems B, Entian KD (2000) The essential protein Fap7 is involved in the oxidative stress response of Saccharomyces cerevisiae. Mol Microbiol 35:936–948CrossRefGoogle Scholar
  6. Keller R (2004) The computer aided resonance tutorial. CANTINA Verlag, GoldauGoogle Scholar
  7. Lienhard GE, Secemski II (1973) P1, P5-di(adenosine-5′)pentaphosphate, a potent multisubstrate inhibitor of adenylate kinase. J Biol Chem 248:1121–1123Google Scholar
  8. Muchmore DC, McIntosh LP, Russell CB, Anderson DE, Dahlquist FW (1989) Expression and nitrogen-15 labeling of proteins for proton and nitrogen-15 nuclear magnetic resonance. Methods Enzymol 177:44–73CrossRefGoogle Scholar
  9. Mulder FAA, Schipper D, Bott R, Boelens R (1999) Altered flexibility in the substrate-binding site of related native and engineered high-alkaline Bacillus subtilisins. J Mol Biol 292:111–123CrossRefGoogle Scholar
  10. Salzmann M, Pervushin K, Wider G, Senn H, Wüthrich K (1998) TROSY in triple-resonance experiments: new perspectives for sequential NMR assignment of large proteins. Proc Natl Acad Sci USA 95:13585–13590ADSCrossRefGoogle Scholar
  11. Walker JE, Saraste M, Runswick MJ, Gay NJ (1982) Distantly related sequences in the alpha- and beta-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold. EMBO J 1:945–951Google Scholar
  12. Wang C, Karpowich N, Hunt JF, Rance M, Palmer AG (2004) Dynamics of ATP-binding cassette contribute to allosteric control, nucleotide binding and energy transduction in ABC transporters. J Mol Biol 342:525–537CrossRefGoogle Scholar

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© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  1. 1.Institut für Molekulare BiowissenschaftenJohann-Wolfgang-Goethe-Universität Frankfurt/M.FrankfurtGermany
  2. 2.Center of Biomolecular Magnetic Resonance (BMRZ)Johann-Wolfgang-Goethe-Universität Frankfurt/M.FrankfurtGermany
  3. 3.Cluster of Excellence „Macromolecular Complexes“Johann-Wolfgang-Goethe-Universität Frankfurt/M.FrankfurtGermany

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