| A tandem affinity purification-based technology platform to study the cell cycle interactome in Arabidopsis thaliana. | |
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MedLine Citation:
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PMID: 17426018 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Defining protein complexes is critical to virtually all aspects of cell biology because many cellular processes are regulated by stable protein complexes, and their identification often provides insights into their function. We describe the development and application of a high throughput tandem affinity purification/mass spectrometry platform for cell suspension cultures to analyze cell cycle-related protein complexes in Arabidopsis thaliana. Elucidation of this protein-protein interaction network is essential to fully understand the functional differences between the highly redundant cyclin-dependent kinase/cyclin modules, which are generally accepted to play a central role in cell cycle control, in all eukaryotes. Cell suspension cultures were chosen because they provide an unlimited supply of protein extracts of actively dividing and undifferentiated cells, which is crucial for a systematic study of the cell cycle interactome in the absence of plant development. Here we report the mapping of a protein interaction network around six known core cell cycle proteins by an integrated approach comprising generic Gateway-based vectors with high cloning flexibility, the fast generation of transgenic suspension cultures, tandem affinity purification adapted for plant cells, matrix-assisted laser desorption ionization tandem mass spectrometry, data analysis, and functional assays. We identified 28 new molecular associations and confirmed 14 previously described interactions. This systemic approach provides new insights into the basic cell cycle control mechanisms and is generally applicable to other pathways in plants. |
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Authors:
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Jelle Van Leene; Hilde Stals; Dominique Eeckhout; Geert Persiau; Eveline Van De Slijke; Gert Van Isterdael; Annelies De Clercq; Eric Bonnet; Kris Laukens; Noor Remmerie; Kim Henderickx; Thomas De Vijlder; Azmi Abdelkrim; Anne Pharazyn; Harry Van Onckelen; Dirk Inzé; Erwin Witters; Geert De Jaeger |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2007-04-09 |
Journal Detail:
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Title: Molecular & cellular proteomics : MCP Volume: 6 ISSN: 1535-9476 ISO Abbreviation: Mol. Cell Proteomics Publication Date: 2007 Jul |
Date Detail:
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Created Date: 2007-07-09 Completed Date: 2007-08-28 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101125647 Medline TA: Mol Cell Proteomics Country: United States |
Other Details:
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Languages: eng Pagination: 1226-38 Citation Subset: IM |
Affiliation:
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Department of Plant Systems Biology, Flanders Institute for Biotechnology, B-9052 Gent, Belgium. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Arabidopsis
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cytology,
metabolism* Arabidopsis Proteins / genetics, metabolism* Cell Cycle / physiology* Cell Cycle Proteins / genetics, metabolism* Cells, Cultured Chromatography, Gel / methods Cloning, Molecular / methods Protein Interaction Mapping Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods |
| Chemical | |
Reg. No./Substance:
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0/Arabidopsis Proteins; 0/Cell Cycle Proteins |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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