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Backbone resonance assignments of the 48 kDa dimeric putative 18S rRNA-methyltransferase Nep1 from Methanocaldococcus jannaschii

  • Jan Philip Wurm
  • Elke Duchardt
  • Britta Meyer
  • Belinda Z. Leal
  • Peter Kötter
  • Karl-Dieter Entian
  • Jens WöhnertEmail author
Article

Abstract

Nep1 from Methanocaldococcus jannaschii is a 48 kDa dimeric protein belonging to the SPOUT-class of S-adenosylmethionine dependent RNA-methyltransferases and acting as a ribosome assembly factor. Mutations in the human homolog are the cause of Bowen-Conradi syndrome. We report here 1H, 15N and 13C chemical shift assignments for the backbone of the protein in its apo state.

Keywords

NMR-assignments Triple resonance experiments RNA-methyltransferase SPOUT-class Nep1 Bowen-Conradi syndrome 

Notes

Acknowledgments

We are grateful to Drs. Andrew P. Hinck and Christian Richter for helpful discussions. This project was supported by start-up funds from the Department of Biochemistry, University of Texas Health Science Center San Antonio (to J.W.), an Aventis Foundation Professorship (to J.W.), the Center of Biomolecular Magnetic Resonance, Johann-Wolfgang-Goethe-University Frankfurt, and the Deutsche Forschungsgemeinschaft (DFG) through the SFB 579 ‘RNA-Ligand Interactions’ (to J.W. and K.-D.E.). The Center of Biomolecular NMR Spectroscopy at UTHSCSA is supported by NIH Grants RR13879 and CA54174.

References

  1. Armistead J, Khatkar S, Meyer B, Mark BL, Patel N, Coghlan G, Lamont RE, Liu S, Wiechert J, Cattini PA, Koetter P, Wrogemann K, Greenberg CR, Entian KD, Zelinski T, Triggs-Raine B (2009) Mutation of a gene essential for ribosome biogenesis, EMG1, causes Bowen-Conradi syndrome. Am J Hum Genet 84:728–739CrossRefGoogle Scholar
  2. Buchhaupt M, Meyer B, Kötter P, Entian KD (2006) Genetic evidence for 18S rRNA binding and an Rps19p assembly function of yeast nucleolar protein Nep1p. Mol Genet Genomics 276:273–284CrossRefGoogle Scholar
  3. Eghbalnia HR, Wang L, Bahrami A, Assadi A, Markley JL (2005) Protein energetic conformational analysis from NMR chemical shifts (PECAN) and its use in determining secondary structural elements. J Biomol NMR 32:71–81CrossRefGoogle Scholar
  4. Eschrich D, Buchhaupt M, Kötter P, Entian KD (2002) Nep1p (Emg1p), a novel protein conserved in eukaryotes and archaea, is involved in ribosome biogenesis. Curr Genet 40:326–338CrossRefGoogle Scholar
  5. Goodsell DS, Olson AJ (2000) Structural symmetry and protein function. Ann Rev Biophys Biomol Struct 29:105–153CrossRefGoogle Scholar
  6. Keller R (2004) The computer aided resonance assignment tutorial. CANTINA Verlag, GoldauGoogle Scholar
  7. Leulliot N, Bohnsack MT, Graille M, Tollervey D, Van Tilbeurgh H (2008) The yeast ribosome synthesis factor Emg1 is a novel member of the superfamily of alpha/beta knot fold methyltransferases. Nucleic Acids Res 36:629–639CrossRefGoogle 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. Taylor AB, Meyer B, Leal BZ, Kötter P, Schirf V, Demeler B, Hart PJ, Entian KD, Wöhnert J (2008) The crystal structure of Nep1 reveals an extended SPOUT-class methyltransferase fold and a pre-organized SAM-binding site. Nucleic Acids Res 36:1542–1554CrossRefGoogle Scholar
  10. Tkaczuk KL, Dunin-Horkawicz S, Purta E, Bujnicki JM (2007) Structural and evolutionary bioinformatics of the SPOUT superfamily of methyltransferases. BMC Bioinformatics 8:73CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Jan Philip Wurm
    • 1
  • Elke Duchardt
    • 1
  • Britta Meyer
    • 1
  • Belinda Z. Leal
    • 3
  • Peter Kötter
    • 1
  • Karl-Dieter Entian
    • 1
  • Jens Wöhnert
    • 1
    • 2
    Email author
  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.Department of BiochemistryThe University of Texas Health Science Center San AntonioSan AntonioUSA

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