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Content archived on 2024-05-29

Plant Anaphase Promoting Complexes: unrevealing APC functions by APC substrates and activators

Objective

Controlled degradation of unwanted and misfolded proteins via the ubiquitin-dependent proteasome system (UPS) is a key regulatory mechanism in eukaryotes. UPS is in the forefront in biological studies, which is essential in all molecular and cellular proce sses. By the UPS, target proteins become polyubiquitinated by consecutive and repeated action of the E1 ubiquitin-activating, E2 ubiquitin-conjugating and E3 the ubiquitin ligase enzymes and then degraded by the 26S proteasome. The substrate specificity of the polyubiquitination is conferred by different E3 enzymes. The anaphase-promoting complex (APC) is one of the most extensively studied E3 ubiquitin ligases in human and yeasts. It is a major regulator of the cell cycle acting from the metaphase to S-pha se and required also for endoreduplication cycles forming polyploid somatic cells. The role of APC is extensively studied in human and yeast, however, in plants the discovery of APC functions has started recently. The proposed project focuses on the functi on of Arabidopsis APCs by using the APC activators to monitor APC activities at organism level and to identify the APC substrates. Unlike single gene copies in yeast or human, A. thaliana contains five CDC20 and three CDH1-type genes; CCS52A1, CCS52A2 and CCS52B. The expression pattern of the CDC20 genes will be tested in transgenic Arabidopsis plants and in synchronised Arabidopsis cell cultures contain CDC20 promoter-reporter gene fusions. The interaction of the CDC20 and CCS52 proteins with the APC will be tested by in vitro and in vivo binding assays and in planta in overexpressing lines. Redundancy of protein functions will be investigated by analysis of different knockout mutants and complementation of the mutants with the other genes. Yeast two hybrid screens will study the overlap and specificity of the CCS52 and CDC20 interacting proteins.

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Keywords

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Topic(s)

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Call for proposal

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FP6-2004-MOBILITY-5
See other projects for this call

Funding Scheme

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EIF - Marie Curie actions-Intra-European Fellowships

Coordinator

CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE
EU contribution
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Total cost

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