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Ubiquitin-dependent proteolysis of KNL2 driven by APC/CCDC20 is critical for centromere integrity and mitotic fidelity

Title: Ubiquitin-dependent proteolysis of KNL2 driven by APC/CCDC20 is critical for centromere integrity and mitotic fidelity
Authors: Kalidass, Manikandan; Jarubula, Venkata Ganesh; Ratnikava, Maryia; Chandra, Jothipriya Ramakrishnan; Le Goff, Samuel; Probst, Aline; Esposito, Silvia; Grasser, Klaus; Bruckmann, Astrid; Gagneux, Jérôme; Prosée, Reinier; Rutten, Twan; Schubert, Veit; Demidov, Dmitri; Lechner, Esther; Steiner, Florian; Genschik, Pascal; Lermontova, Inna
Contributors: Leibniz Institute of Plant Genetics and Crop Plant Research Gatersleben (IPK-Gatersleben); Université Clermont Auvergne (UCA); Universität Regensburg = University of Regensburg = Université de Ratisbonne Allemagne (UR); Université de Genève = University of Geneva (UNIGE); Institut de Biologie Moléculaire des Plantes (IBMP); Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS); Deutsche Forschungsgemeinschaft (DFG) grants (DFG LE 2299/5-1 and DFG LE 2299/8-1); SFB960-A6 to K.D.G.; COST Action, CA16212; Groupement de Recherche EPIPLANT (GDR EPIPLANT); Swiss National Science Foundation grants (31003A_175606 and 310030_197762); ANR-11-JSV2-0009,Dynam'Het,Organisation dynamique de l'hétérochromatine au cours du développement chez Arabidopsis thaliana(2011); ANR-10-IDEX-0002,UNISTRA,Par-delà les frontières, l'Université de Strasbourg(2010)
Source: ISSN: 1040-4651.
Publisher Information: CCSD; American Society of Plant Biologists (ASPB)
Publication Year: 2025
Collection: HAL Clermont Auvergne (Université Blaise Pascal Clermont-Ferrand / Université d'Auvergne)
Subject Terms: [SDV]Life Sciences [q-bio]
Description: International audience ; Kinetochores are large protein complexes that serve as attachment sites for spindle microtubules, ensuring proper chromosome segregation during cell division. KINETOCHORE NULL2 (αKNL2) is a key kinetochore protein required for the incorporation of the centromeric histone variant CENH3. The precise regulation of αKNL2 levels is crucial, but the molecular mechanisms controlling this process remain largely unexplored. In this study, we demonstrated that the Anaphase-Promoting Complex/Cyclosome (APC/C) mediates the ubiquitin-dependent proteolysis of αKNL2 during mitosis. Our findings revealed that αKNL2 accumulates in the presence of 26S proteasome inhibitors, and our yeast 2-hybrid and proteomic screens showed that proteins from the ubiquitin-proteasome pathway interact with KNL2 in Arabidopsis (Arabidopsis thaliana) and nematode (Caenorhabditis elegans). Arabidopsis αKNL2 directly interacts with Anaphase-Promoting Complex subunit 10 (APC10) and Cell Division Cycle 20.1 (CDC20.1), 2 substrate recognition components of the APC/C. RNAi-mediated depletion of APC/C resulted in the accumulation and mislocalization of endogenous αKNL2. Additionally, mutation or deletion of the D-box1 region, or substitution of residues K336 and K339, impaired αKNL2 degradation. The expression of a proteasome-resistant αKNL2 variant in planta caused severe defects in growth, fertility, and mitotic division. These findings show that APC/CCDC20-mediated degradation of αKNL2 is critical for proper kinetochore function and centromere integrity.
Document Type: article in journal/newspaper
Language: English
DOI: 10.1093/plcell/koaf164
Availability: https://hal.science/hal-05363668; https://hal.science/hal-05363668v1/document; https://hal.science/hal-05363668v1/file/koaf164.pdf; https://doi.org/10.1093/plcell/koaf164
Rights: https://creativecommons.org/licenses/by/4.0/ ; info:eu-repo/semantics/OpenAccess
Accession Number: edsbas.6B819EA7
Database: BASE