Targeted disruption of linkage-specific ubiquitylation reveals a key role of K29-linked ubiquitylation in epigenome integrity

Arroyo-Gomez, J.; Wang, J.; Guerillon, C.; Isaakova, E.; Reveron-Gomez, N.; Koutrouli, M.; Rahmanto, A. S.; Mitrofanov, K.; Ingham, A.; Schovsbo, S.; Weischenfeldt, K.; Coscia, F.; Typas, D.; Volker-Albert, M.; Solis, V.; Juhl Jensen, L.; Groth, A.; Mund, A.; Beli, P.; Shearer, R. F.; Mailand, N.

doi:10.1101/2024.10.29.620783

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bioRxiv · 10.1101/2024.10.29.620783

Targeted disruption of linkage-specific ubiquitylation reveals a key role of K29-linked ubiquitylation in epigenome integrity

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Abstract

Linkage-specific ubiquitin chains dictate the functional outcome of numerous critical ubiquitin-dependent signaling processes. However, the functions and targets of several poly-ubiquitin topologies remain poorly defined due to a paucity of tools for their specific detection and manipulation. To remedy this knowledge gap, we applied a cell-based ubiquitin replacement strategy enabling targeted conditional abrogation of each of the seven lysine-based ubiquitin chain types in human cells to profile system-wide impacts of disabling formation of individual chain types. Focusing on K29-linked ubiquitylation, we reveal a strong association of this linkage type with chromatin-associated proteins and show that the H3K9me3 methyltransferase SUV39H1 is a prominent cellular target of this modification. We demonstrate that K29-linked ubiquitylation is essential for proteasomal degradation of SUV39H1 despite its extensive modification by K48-linked ubiquitylation, and that K29-linked ubiquitylation of SUV39H1 is catalyzed and reversed by TRIP12 and TRABID, respectively. Preventing K29-linked ubiquitylation-mediated control of SUV39H1 stability deregulates the H3K9me3 landscape, but not other histone marks. Collectively, our ubiquitin replacement cell line panel and datasets provide valuable resources for illuminating cellular functions of linkage-specific ubiquitin chains and establish a key role of K29-linked ubiquitylation in epigenome integrity.

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Arroyo-Gomez, J., Wang, J., Guerillon, C., Isaakova, E., Reveron-Gomez, N., Koutrouli, M., Rahmanto, A. S., Mitrofanov, K., Ingham, A., Schovsbo, S., Weischenfeldt, K., Coscia, F., Typas, D., Volker-Albert, M., Solis, V., Juhl Jensen, L., Groth, A., Mund, A., Beli, P., Shearer, R. F., Mailand, N.. 2024-11-02. Targeted disruption of linkage-specific ubiquitylation reveals a key role of K29-linked ubiquitylation in epigenome integrity. https://doi.org/10.1101/2024.10.29.620783

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