Biochemical, biophysical, and functional characterisation of the E3 ubiquitin ligase APC/C regulator CDC20 from Arabidopsis thaliana

• DOI: 10.3389/fphys.2022.938688
• Publication Date: 2022-07-25T07:04:49Z
• Authors: Maria-Alexa Cosma; Natalie L. Curtis; Charlotte Pain; Verena Kriechbaumer; Victor M. Bolanos...
• Keywords: Degron; CDC20; Cullin; PLK1; Spindle checkpoint; Anaphase-promoting complex

The Anaphase Promoting Complex (APC/C), a large cullin-RING E3-type ubiquitin ligase, constitutes the ultimate target of Spindle Assembly Checkpoint (SAC), an intricate regulatory circuit that ensures high fidelity chromosome segregation in eukaryotic organisms by delaying onset anaphase until each is properly bi-oriented on mitotic spindle. Cell-division cycle protein 20 homologue (CDC20) key regulator APC/C function mitosis. formation APC/CCDC20 complex required for ubiquitination and degradation select substrates, which necessary to maintain state. In contrast roles CDC20 animal species, little known about regulation plants. Here we address this gap knowledge report expression insect cells; biochemical biophysical characterisation Arabidopsis thaliana (AtCDC20) WD40 domain; nuclear cytoplasmic distribution full-length AtCDC20 when transiently expressed tobacco We also show most degrons share sequence similarity other eukaryotes, arguing favour conserved degron functions AtCDC20. However, important exceptions were noted such as lack canonical MAD1 binding motif; fully RRY-box all six isoforms instead CRY-box motif, low conservation residues be phosphorylated BUB1 PLK1 species ensure robust SAC response. Taken together, our studies provide insights into structure evolution signalling