Revisiting degron motifs in human AURKA required for its targeting by APC/C<sup>FZR1</sup>

Abdelbaki, Ahmed; Ascanelli, Camilla; Okoye, Cynthia N; Akman, Hesna Begüm; Janson, Giacomo; Min, Mingwei; Marcozzi, Chiara; Hagting, Anja; Grant, Rhys; Luca, Maria De; Asteriti, Italia Anna; Guarguaglini, Giulia; Paiardini, Alessandro; Lindon, Catherine

doi:10.26508/lsa.202201372

Cited by 4 publications

(2 citation statements)

References 48 publications

(70 reference statements)

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“…For instance, the spindle-associated protein FAM83D 52 exhibited a significant accumulation in ANAPC4-depleted cells (Figure 6D) and a protein oscillation consistent with the APC/C substrate pattern (Supplementary Figures 7E and 7F). This observation aligns with other examples of APC/C substrates that lack canonical degradation motifs, such as AURKA and SGO1 which contain a non-canonical D box 53,54 . This set of proteins with non-canonical degradation motifs raises intriguing questions about the intricacies underlying their degradation mechanisms.…”

Section: Degradation Mechanisms Of Ccd Proteinssupporting

confidence: 89%

High resolution profiling of cell cycle-dependent protein and phosphorylation abundance changes in non-transformed cells

Rega,

Tsitsa,

Roumeliotis

et al. 2024

Preprint

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The cell cycle governs a precise series of molecular events, regulated by coordinated changes in protein and phosphorylation abundance, that culminates in the generation of two daughter cells. Here, we present a proteomic and phosphoproteomic analysis of the human cell cycle in hTERT-RPE-1 cells using deep quantitative mass spectrometry by isobaric labelling. Through analysing non-transformed cells, and improving the temporal resolution and coverage of key cell cycle regulators, we present a dataset of cell cycle-dependent protein and phosphorylation site oscillation that offers a foundational reference for investigating cell cycle regulation. These data reveal uncharacterised regulatory intricacies including proteins and phosphorylation sites exhibiting previously unreported cell cycle-dependent oscillation, and novel proteins targeted for degradation during mitotic exit. Integrated with complementary resources, our data link cycle-dependent abundance dynamics to functional changes and are accessible through the Cell Cycle database (CCdb), an interactive web-based resource for the cell cycle community.

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Section: Degradation Mechanisms Of Ccd Proteinssupporting

confidence: 89%

High resolution profiling of cell cycle-dependent protein and phosphorylation abundance changes in non-transformed cells

Rega,

Tsitsa,

Roumeliotis

et al. 2024

Preprint

View full text Add to dashboard Cite

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“…8A). The D-box sequences were swapped into an RxxL motif previously shown to have no degron activity (Abdelbaki et al 2022) which we here refer to as ‘D0’, adjacent to the endogenous C-terminal IDR of AURKA to enable processing of the ubiquitinated fusion proteins at the 26S proteasome. We found that all four new D-box variants tested could target mNeon for degradation, with timing consistent with targeting by APC/C Cdc20 (Fig.…”

Section: Resultsmentioning

confidence: 99%

Development of D-box peptides to inhibit the Anaphase Promoting Complex/Cyclosome

Eapen,

Okoye,

Stubbs

et al. 2024

Preprint

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E3 ubiquitin ligases engage their substrates via 'degrons' - short linear motifs typically located within intrinsically disordered regions of substrates. As these enzymes are large, multi-subunit complexes that generally lack natural small-molecule ligands and are hard to drug via conventional means, alternative strategies are needed to target them in diseases, and peptide-based inhibitors derived from degrons represent a promising approach. Here we explore peptide inhibitors of Cdc20, a substrate-recognition subunit and activator of the E3 ubiquitin ligase the anaphase promoting complex/cyclosome (APC/C) that is essential in mitosis and consequently of interest as an anti-cancer target. APC/C engages substrates via degrons that include the 'Destruction box' (D-box) motif. We used a rational design approach to construct binders containing unnatural amino acids aimed at better filling a hydrophobic pocket on the surface of Cdc20. We confirmed binding by thermal-shift assays and surface plasmon resonance and determined the structures of a number of the Cdc20-peptide complexes. Using a cellular thermal shift assay we confirmed that the D-box peptides also bind to and stabilise Cdc20 in the cell. We found that the D-box peptides inhibit ubiquitination activity of APC/CCdc20 and are more potent than the small molecule inhibitor Apcin. Lastly, these peptides function as portable degrons capable of driving the degradation of a fused fluorescent protein. Interestingly, we find that although inhibitory activity of the peptides correlates with Cdc20-binding affinity, degradation efficacy does not, which may be due to the complex nature of APC/C regulation and effects of degron binding of subunit recruitment and conformational changes. Our study lays the groundwork for the further development of these peptides as molecular therapeutics for blocking APC/C as well as potentially also for harnessing APC/C for targeted protein degradation.

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Single nucleus transcriptomics of ventral midbrain identifies glial activation associated with chronic opioid use disorder

Wei,

Lambert,

Valada

et al. 2023

Nat Commun

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Dynamic interactions of neurons and glia in the ventral midbrain mediate reward and addiction behavior. We studied gene expression in 212,713 ventral midbrain single nuclei from 95 individuals with history of opioid misuse, and individuals without drug exposure. Chronic exposure to opioids was not associated with change in proportions of glial and neuronal subtypes, however glial transcriptomes were broadly altered, involving 9.5 − 6.2% of expressed genes within microglia, oligodendrocytes, and astrocytes. Genes associated with activation of the immune response including interferon, NFkB signaling, and cell motility pathways were upregulated, contrasting with down-regulated expression of synaptic signaling and plasticity genes in ventral midbrain non-dopaminergic neurons. Ventral midbrain transcriptomic reprogramming in the context of chronic opioid exposure included 325 genes that previous genome-wide studies had linked to risk of substance use traits in the broader population, thereby pointing to heritable risk architectures in the genomic organization of the brain’s reward circuitry.

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Revisiting degron motifs in human AURKA required for its targeting by APC/C^FZR1

Cited by 4 publications

References 48 publications

High resolution profiling of cell cycle-dependent protein and phosphorylation abundance changes in non-transformed cells

High resolution profiling of cell cycle-dependent protein and phosphorylation abundance changes in non-transformed cells

Development of D-box peptides to inhibit the Anaphase Promoting Complex/Cyclosome

Single nucleus transcriptomics of ventral midbrain identifies glial activation associated with chronic opioid use disorder

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Revisiting degron motifs in human AURKA required for its targeting by APC/CFZR1

Cited by 4 publications

References 48 publications

High resolution profiling of cell cycle-dependent protein and phosphorylation abundance changes in non-transformed cells

High resolution profiling of cell cycle-dependent protein and phosphorylation abundance changes in non-transformed cells

Development of D-box peptides to inhibit the Anaphase Promoting Complex/Cyclosome

Single nucleus transcriptomics of ventral midbrain identifies glial activation associated with chronic opioid use disorder

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Revisiting degron motifs in human AURKA required for its targeting by APC/C^FZR1