Quantitative phosphoproteomics revealswidespread full phosphorylation site occupancy during mitosis Journal Article
| Authors: | Olsen, J. V.; Vermeulen, M.; Santamaria, A.; Kumar, C.; Miller, M. L.; Jensen, L. J.; Gnad, F.; Cox, J.; Jensen, T. S.; Nigg, E. A.; Brunak, S.; Mann, M. |
| Article Title: | Quantitative phosphoproteomics revealswidespread full phosphorylation site occupancy during mitosis |
| Abstract: | Eukaryotic cells replicate by a complex series of evolutionarily conserved events that are tightly regulated at defined stages of the cell division cycle. Progression through this cycle involves a large number of dedicated protein complexes and signaling pathways, and deregulation of this process is implicated in tumorigenesis. We applied high-resolution mass spectrometry-based proteomics to investigate the proteome and phosphoproteome of the human cell cycle on a global scale and quantified 6027 proteins and 20,443 unique phosphorylation sites and their dynamics. Co-regulated proteins and phosphorylation sites were grouped according to their cell cycle kinetics and compared to publicly available messenger RNA microarray data. Most detected phosphorylation sites and more than 20% of all quantified proteins showed substantial regulation, mainly in mitotic cells. Kinase-motif analysis revealed global activation during S phase of the DNA damage response network, which was mediated by phosphorylation by ATM or AT or DNA-dependent protein kinases. We determined site-specific stoichiometry of more than 5000 sites and found that most of the up-regulated sites phosphorylated by cyclin-dependent kinase 1 (CDK1) or CDK2 were almost fully phosphorylated in mitotic cells. In particular, nuclear proteins and proteins involved in regulating metabolic processes have high phosphorylation site occupancy in mitosis. This suggests that these proteins may be inactivated by phosphorylation in mitotic cells. Copyright 2008 by the American Association for the Advancement of Science; all rights reserved. |
| Keywords: | signal transduction; controlled study; protein phosphorylation; protein array analysis; human cell; genetics; flow cytometry; methodology; mitosis; mass spectrometry; proteome; metabolism; dna damage; cell cycle; cell cycle s phase; cluster analysis; gene expression profiling; protein degradation; protein binding; hela cell; hela cells; phosphorylation; proteomics; physiology; amino acid sequence; cell culture; messenger rna; eukaryota; oligonucleotide array sequence analysis; quantitative analysis; substrate specificity; immunoblotting; atm protein; phosphoproteins; binding site; binding sites; upregulation; dna microarray; enzyme specificity; cell cycle regulation; atr protein; protein microarray; phosphoprotein; cyclin dependent kinase 1; stoichiometry; cyclin dependent kinase 2; cyclin-dependent kinase 2; deregulation; dna dependent protein kinase; cdk2 protein, human; cdc2 protein kinase |
| Journal Title: | Science Signaling |
| Volume: | 3 |
| Issue: | 104 |
| ISSN: | 1945-0877 |
| Publisher: | American Association for the Advancement of Science |
| Date Published: | 2010-01-12 |
| Start Page: | ra3 |
| Language: | English |
| DOI: | 10.1126/scisignal.2000475 |
| PUBMED: | 20068231 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 54" - "Export Date: 20 April 2011" - "Source: Scopus" |
