4.5 Article

Shotgun proteomic analysis of the unicellular alga Ostreococcus tauri

Journal

JOURNAL OF PROTEOMICS
Volume 74, Issue 10, Pages 2060-2070

Publisher

ELSEVIER
DOI: 10.1016/j.jprot.2011.05.028

Keywords

Ostreococcus tauri; Algal proteomics; Label-free quantitation; N-15 metabolic labeling; Circadian rhythms; Phosphopeptide enrichment

Funding

  1. Centre for Systems Biology at Edinburgh (CSBE) which is a Centre for Integrative Systems Biology (CISB)
  2. BBSRC
  3. EPSRC [BB/D019621/1]
  4. BBSRC [BB/D019621/1, BB/F005466/1] Funding Source: UKRI
  5. Biotechnology and Biological Sciences Research Council [BB/D019621/1, BB/F005466/1] Funding Source: researchfish

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Ostreococcus tauri is a unicellular green alga and amongst the smallest and simplest free-living eukaryotes. The O. tauri genome sequence was determined in 2006. Molecular, physiological and taxonomic data that has been generated since then highlight its potential as a simple model species for algae and plants. However, its proteome remains largely unexplored. This paper describes the global proteomic study of O. tauri, using mass spectrometry-based approaches: phosphopeptide enrichment, cellular fractionation, label-free quantification and N-15 metabolic labeling. The O. tauri proteome was analyzed under the following conditions: sampling at different times during the circadian cycle, after 24 h of illumination, after 24 h of darkness and under various nitrogen source supply levels. Cell cycle related proteins such as dynamin and kinesin were significantly up-regulated during the daylight-to-darkness transition. This is reflected by their higher intensity at ZT13 and this transition phase coincides with the end of mitosis. Proteins involved in several metabolic mechanisms were found to be up-regulated under low nitrogen conditions, including carbon storage pathways, glycolysis, phosphate transport, and the synthesis of inorganic polyphosphates. Ostreococcus tauri responds to low nitrogen conditions by reducing its nitrogen assimilation machinery which suggests an atypical adaptation mechanism for coping with a nutrient-limited environment. (C) 2011 Elsevier B.V. All rights reserved.

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