Proteomics Analysis of the Nucleolus in Adenovirus-infected Cells

Yun W. Lam, Vanessa C. Evans, Kate J. Heesom, Angus I. Lamond, David A. Matthews

Research output: Contribution to journalArticlepeer-review

101 Citations (Scopus)

Abstract

Adenoviruses replicate primarily in the host cell nucleus, and it is well established that adenovirus infection affects the structure and function of host cell nucleoli in addition to coding for a number of nucleolar targeted viral proteins. Here we used unbiased proteomics methods, including high throughput mass spectrometry coupled with stable isotope labeling by amino acids in cell culture (SILAC) and traditional two-dimensional gel electrophoresis, to identify quantitative changes in the protein composition of the nucleolus during adenovirus infection. Two-dimensional gel analysis revealed changes in six proteins. By contrast, SILAC-based approaches identified 351 proteins with 24 proteins showing at least a 2-fold change after infection. Of those, four were previously reported to have aberrant localization and/or functional relevance during adenovirus infection. In total, 15 proteins identified as changing in amount by proteomics methods were examined in infected cells using confocal microscopy. Eleven of these proteins showed altered patterns of localization in adenovirus-infected cells. Comparing our data with the effects of actinomycin D on the nucleolar proteome revealed that adenovirus infection apparently specifically targets a relatively small subset of nucleolar antigens at the time point examined.

Original languageEnglish
Pages (from-to)117-130
Number of pages14
JournalMolecular and Cellular Proteomics
Volume9
Issue number1
Early online date7 Oct 2009
DOIs
Publication statusPublished - 1 Jan 2010
Externally publishedYes

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