%0 Journal Article %T Sensitive detection of protein ubiquitylation using a protein-fragment complementation assay %+ Institut de Génétique et Développement de Rennes (IGDR) %+ Institut Jacques Monod (IJM (UMR_7592)) %A Le Boulch, Marie %A Brossard, Audrey %A Le Dez, Gaëlle %A Léon, Sébastien %A Rabut, Gwenaël %Z PICS07394, Centre National de la Recherche Scientifique %Z Institut National de la Santé et de la Recherche Médicale %Z Université de Rennes 1 %Z ANR-16-CE13-0002-02, Agence Nationale de la Recherche %Z Ligue Contre le Cancer %Z Biosit %< avec comité de lecture %@ 0021-9533 %J Journal of Cell Science %I Company of Biologists %V 133 %N 12 %P jcs240093 %8 2020-06-15 %D 2020 %R 10.1242/jcs.240093 %M 32409563 %K Ubiquitin %K NanoBiT %K NanoLuc %K PCA %K Ubiquitination %K UbiCREST %Z Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC] %Z Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM]Journal articles %X Ubiquitylation is a reversible post-translational protein modification that regulates a multitude of cellular processes. Detection of ubiquitylated proteins is often challenging, because of their low abundance. Here, we present NUbiCA, a sensitive protein-fragment complementation assay to facilitate the monitoring of ubiquitylation events in cultured cells and model organisms. Using yeast as a model system, we demonstrate that NUbiCA enables to accurately monitor mono- and poly-ubiquitylation of proteins expressed at endogenous levels. We also show that it can be applied to decipher the topology of ubiquitin conjugates. Moreover, we assembled a genome wide collection of yeast strains ready to investigate the ubiquitylation of proteins with this new assay. This resource will facilitate the analysis of local or transient ubiquitylation events that are difficult to detect with current methods. %G English %2 https://univ-rennes.hal.science/hal-02635146/document %2 https://univ-rennes.hal.science/hal-02635146/file/Le%20Boulch%20et%20al-2020-Sensitive%20detection%20of%20protein%20ubiquitylation.pdf %L hal-02635146 %U https://univ-rennes.hal.science/hal-02635146 %~ UNIV-RENNES1 %~ CNRS %~ IJM %~ IFR140 %~ IGDR %~ STATS-UR1 %~ UR1-UFR-SVE %~ UR1-HAL %~ UR1-SDV %~ IGDR-UP %~ TEST-UR-CSS %~ UNIV-RENNES %~ UNIV-PARIS %~ UNIVERSITE-PARIS %~ UP-SCIENCES %~ TEST-HALCNRS %~ ANR %~ UR1-BIO-SA