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Zn-binding AZUL domain of human ubiquitin protein ligase Ube3A

Journal of Biomolecular NMR volume 51, Article number: 185 (2011) Cite this article

Abstract

Ube3A (also referred to as E6AP for E6 Associated Protein) is a E3 ubiquitin-protein ligase implicated in the development of Angelman syndrome by controlling degradation of synaptic protein Arc and oncogenic papilloma virus infection by controlling degradation of p53. This article describe the solution NMR structure of the conserved N-terminal domain of human Ube3A (residues 24-87) that contains two residues (Cys44 and Arg62) found to be mutated in patients with Angelman syndrome. The structure of this domain adopts a novel Zn-binding fold we called AZUL (Amino-terminal Zn-finger of Ube3a Ligase). The AZUL domain has a helix-loop-helix architecture with a Zn ion coordinated by four Cys residues arranged in Cys-X4-Cys-X4-Cys-X28-Cys motif. Three of the Zn-bound residues are located in a 23-residue long and well structured loop that connects two α-helicies.

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References

  • Alberts IL, Nadassy K, Wodak SJ (1998) Analysis of zinc binding sites in protein crystal structures. Protein Sci 7(8):1700–1716

    Article  Google Scholar 

  • Bai JL, Qu YJ et al (2011) A novel missense mutation of the ubiquitin protein ligase E3A gene in a patient with Angelman syndrome. Chin Med J Engl 124(1):84–88

    Google Scholar 

  • Beaudenon S, Huibregtse JM (2008) HPV E6, E6AP and cervical cancer. BMC Biochem 9 suppl 1:S4.

  • Bhattacharya A, Tejero R, Montelione GT (2007) Evaluating protein structures determined by structural genomics consortia. Proteins 66(4):778–795

    Article  Google Scholar 

  • Brünger AT, Adams PD et al (1998) Crystallography & NMR system: A new software suite for macromolecular structure determination. Acta Crystallogr D Biol Crystallogr 54(Pt 5):905–921

    Article  Google Scholar 

  • Cooper EM, Hudson AW et al (2004) Biochemical analysis of Angelman syndrome-associated mutations in the E3 ubiquitin ligase E6-associated protein. J Biol Chem 279(39):41208–41217

    Article  Google Scholar 

  • Delaglio F, Grzesiek S et al (1995) Nmrpipe—a multidimensional spectral processing system based on unix Pipes. J Biomol NMR 6(3):277–293

    Article  Google Scholar 

  • Goddard TD, Kneller DG (2003) Sparky—NMR assignment and integration software.

  • Greer PL, Hanayama R et al (2010) The Angelman syndrome protein Ube3A regulates synapse development by ubiquitinating arc. Cell 140(5):704–716

    Article  Google Scholar 

  • Güntert P (2004) Automated NMR structure calculation with CYANA. Methods Mol Biol 278:353–378

    Google Scholar 

  • Holm L, Rosenström P (2010) Dali server: conservation mapping in 3D. Nucleic Acids Res 38(Web Server issue): W545–W549

    Google Scholar 

  • Huang YJ, Powers R, Montelione GT (2005) Protein NMR recall, precision, and F-measure scores (RPF scores): structure quality assessment measures based on information retrieval statistics. J Am Chem Soc 127:1665–1674

    Article  Google Scholar 

  • Kay LE (1995) Pulsed field gradient multi-dimensional NMR methods for the study of protein structure and dynamics in solution. Prog Biophys Mol Biol 63(3):277–299

    Article  Google Scholar 

  • Koradi R, Billeter M and Wuthrich K (1996) MOLMOL: a program for display and analysis of macromolecular structures. J Mol Graph 14(1):51–55, 29–32.

    Google Scholar 

  • Kornhaber GJ, Snyder D, Moseley HNB, Montelione GT (2006) Identification of zinc-ligated cysteine residues based on C-13 alpha and C-13 beta chemical shift data. J Biomol NMR 34(4):259–269

    Article  Google Scholar 

  • Lemak A, Gutmanas A et al (2011) A novel strategy for NMR resonance assignment and protein structure determination. J Biomol NMR 49(1):27–38

    Article  Google Scholar 

  • Linge JP, Williams MA et al (2003) Refinement of protein structures in explicit solvent. Proteins 50(3):496–506

    Article  Google Scholar 

  • Malzac P, Webber H et al (1998) Mutation analysis of UBE3A in Angelman syndrome patients. Am J Hum Genet 62(6):1353–1360

    Article  Google Scholar 

  • Shen Y, Lange O et al (2008) Consistent blind protein structure generation from NMR chemical shift data. Proc Natl Acad Sci USA 105(12):4685–4690

    ADS  Article  Google Scholar 

  • Walboomers JM, Jacobs MV et al (1999) Human papillomavirus is a necessary cause of invasive cervical cancer worldwide. J Pathol 189(1):12–19

    Article  Google Scholar 

  • Williams CA, Driscoll DJ, Dagli AI (2010) Clinical and genetic aspects of Angelman syndrome. Genet Med 12(7):385–395

    Article  Google Scholar 

  • Yee A, Chang X et al (2002) An NMR approach to structural proteomics. Proc Natl Acad Sci USA 99(4):1825–1830

    ADS  Article  Google Scholar 

Download references

Acknowledgments

Authors are thankful to Peter Loppnau for Ube3A constructs cloning. This work is supported by the US National Institute of Health Protein Structure Initiative (P50-GM62413-01 and GM67965) through the Northeast Structural and the Structural Genomics Consortium a registered charity (number 1097737) that receives funds from the Canadian Institutes of Health Research, the Canadian Foundation for Innovation, Eli Lilly, Genome Canada through the Ontario Genomics Institute, GlaxoSmithKline, Novartis, the Ontario Minstry of Research and Innovation, Pfizer, and the Wellcome Trust. CHA holds a Canada Research Chair in Structural Proteomics. IB is supported by grant 1P30GM092369 from the NIH.

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Authors and Affiliations

  1. Ontario Cancer Institute, Campbell Family Cancer Research Institute and Department of Medical Biophysics, University of Toronto, and Northeast Structural Genomics Consortium, 101 College Street, Toronto, ON, M5G 1L7, Canada

    Alexander Lemak, Adelinda Yee & Cheryl H. Arrowsmith

  2. Department of Molecular Microbial and Structural Biology, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT, 06030, USA

    Irina Bezsonova

  3. Structural Genomics Consortium, University of Toronto, 101 College Street, Toronto, ON, M5G 1L7, Canada

    Sirano Dhe-Paganon & Cheryl H. Arrowsmith

  4. Department of Physiology, University of Toronto, 100 College Street, Toronto, ON, M5G 1L5, Canada

    Sirano Dhe-Paganon

Authors
  1. Alexander Lemak
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  2. Adelinda Yee
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  3. Irina Bezsonova
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  4. Sirano Dhe-Paganon
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  5. Cheryl H. Arrowsmith
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Corresponding authors

Correspondence to Irina Bezsonova, Sirano Dhe-Paganon or Cheryl H. Arrowsmith.

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Lemak, A., Yee, A., Bezsonova, I. et al. Zn-binding AZUL domain of human ubiquitin protein ligase Ube3A. J Biomol NMR 51, 185 (2011). https://doi.org/10.1007/s10858-011-9552-y

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  • DOI: https://doi.org/10.1007/s10858-011-9552-y

Keywords

  • Ube3A
  • E6-AP
  • E3A ubiquitin ligase
  • AZUL
  • Angelman syndrome
  • Zn-finger
  • HPV
  • NMR