Nucleolar Proteomics

Abstract

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 μg 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

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Further Reading

Housby JN (ed.) (2001) Mass Spectrometry and Genomic Analysis. Dordrecht: Kluwer.

Link AJ (ed.) (1999) 2‐D Proteome Analysis Protocols. Totowa, NJ: Humana Press.

Wilkins MR, Williams KL, Appel RD and Hochstrasser DF (eds.) (1997) Proteome Research: New Frontiers in Functional Genomics. Berlin: Springer‐Verlag.

Web Links

Basic Local Alignment Search Tool (BLAST) http://www.expasy.org

SWISS‐2DPAGE database http://www.expasy.org/ch2d

Protein Info Search engine http://prowl.rockfelle.edu/

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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]