Nucleolar Proteomics

Yohann Couté, INSERM, Faculté de Médecine Lyon‐R.T.H. Laennec, Lyon, France
Alexander Scherl, Geneva University Hospital, Switzerland
Régis Dieckmann, Geneva University Hospital, Switzerland
Aleth Callé, INSERM, Faculté de Médecine Lyon‐R.T.H. Laennec, Lyon, France
Christine Hoogland, Swiss Institute of Bioinformatics, Ganeva, Switzerland
Karine Kindbeiter, INSERM, Faculté de Médecine Lyon‐R.T.H. Laennec, Lyon, France
Catherine Déon, Geneva University Hospital, Switzerland
Anna Greco, INSERM, Faculté de Médecine Lyon‐R.T.H. Laennec, Lyon, France
Jean‐Charles Sanchez, Geneva University Hospital, Switzerland
Jean‐Jacques Diaz, INSERM, Faculté de Médecine Lyon‐R.T.H. Laennec, Lyon, France
Denis Hochstrasser, Geneva University Hospital, Switzerland

Published online: January 2006

DOI: 10.1038/npg.els.0006190

The nucleolus is a dense membrane-free nuclear domain, visible by light microscopy. It is known that the nucleolus is the site of ribosomal gene transcription and ribosome assembly. However, it appears that the nucleolus also plays a key role in many other biological processes.

Keywords: nucleolus; mass spectrometry; proteomics; nucleolar functions; annotated naps

Figure 1. electron microscopy image of a purified nucleolus showing the characteristic ultrastructure: fibrillar center (FC), dense fibrillar component (DFC) and granular component (GC).
Figure 2. (a) Total ion trace; (b) reconstructed ion chromatogram of m/z=957.6 (at 27.4 min, a peptide with this m/z is eluted); (c) MS spectrum of the doubly charged peptide (m/z=957.6); (d) tandem mass spectrum of the fragments of the peptide corresponding to QIIEEDAALVEIGPR. This sequence matched a single protein, similar to RIKEN cDNA 1110064N10 gene (Q96DH1).
Figure 3. SDS-PAGEseparation of 15 g of nucleolar proteins stained with Coomassie blue. This gel lane was sliced into 108 bands. Proteins from each band were digested and separately analyzed by microcapillary LC-MS/MS. One hundred and ninety proteins were identified. In band 12, 15 different proteins were identified.
Figure 4. Functional classification of the 213 identified proteins in human nucleoli. An extensive bibliographic analysis and sequence homology searches allowed the grouping of the identified nucleolar proteins into different functional classes according to the biological processes in which they are known and/or proposed to be involved. The number of proteins belonging to each class is indicated.
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 References
    Andersen JS, Lyon CE, Fox AH, et al. (2002) Direct proteomic analysis of the human nucleolus. Current Biology 12: 1–11.
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 Further Reading
    book Housby JN (ed.) (2001) Mass Spectrometry and Genomic Analysis. Dordrecht: Kluwer.
    book Link AJ (ed.) (1999) 2-D Proteome Analysis Protocols. Totowa, NJ: Humana Press.
    book Wilkins MR, Williams KL, Appel RD and Hochstrasser DF (eds.) (1997) Proteome Research: New Frontiers in Functional Genomics. Berlin: Springer-Verlag.
 Web Links
    ePath Basic Local Alignment Search Tool (BLAST) http://www.expasy.org
    ePath SWISS-2DPAGE database http://www.expasy.org/ch2d
    ePath Protein Info Search engine http://prowl.rockfelle.edu/

Related Sub-Topics:

Basic Cell Biology, Protein, RNA and DNA | Cell Compartments and Cellular Organelles | Techniques in Cell Biology | Protein Structure | Mass Spectrometry | Biochemical and Biophysical Techniques | Proteins: Structure, Function, Metabolism
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Article Title: Nucleolar Proteomics
 
How to Cite close
Couté, Yohann, Scherl, Alexander, Dieckmann, Régis, Callé, Aleth, Hoogland, Christine, Kindbeiter, Karine, Déon, Catherine, Greco, Anna, Sanchez, Jean‐Charles, Diaz, Jean‐Jacques, and Hochstrasser, Denis(Jan 2006) Nucleolar Proteomics. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0006190]