A chemical probe to modulate human GID4 Pro/N-degron interactions

期刊论文
发表日期：

2024年5月21日

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Dominic D. G. Owens,
Matthew E. R. Maitland,
Aliakbar Khalili Yazdi,
Xiaosheng Song,
Viviane Reber,
Martin P. Schwalm,
Raquel A. C. Machado,
Nicolas Bauer,
Xu Wang,
Magdalena M. Szewczyk,
Cheng Dong,
Aiping Dong,
Peter Loppnau,
Matthew F. Calabrese,
Matthew S. Dowling,
Jisun Lee,
Justin I. Montgomery,
Thomas N. O’Connell,
Chakrapani Subramanyam,
Feng Wang,
Ella C. Adamson,
Matthieu Schapira,
Matthias Gstaiger,
Stefan Knapp,
Masoud Vedadi,
Jinrong Min,
Gilles A. Lajoie,
Dalia Barsyte-Lovejoy,
Dafydd R. Owen,
Caroline Schild-Poulter,
Cheryl H. Arrowsmith

摘要

The C-terminal to LisH (CTLH) complex is a ubiquitin ligase complex that recognizes substrates with Pro/N-degrons via its substrate receptor Glucose-Induced Degradation 4 (GID4), but its function and substrates in humans remain unclear. Here, we report PFI-7, a potent, selective and cell-active chemical probe that antagonizes Pro/N-degron binding to human GID4. Use of PFI-7 in proximity-dependent biotinylation and quantitative proteomics enabled the identification of GID4 interactors and GID4-regulated proteins. GID4 interactors are enriched for nucleolar proteins, including the Pro/N-degron-containing RNA helicases DDX21 and DDX50. We also identified a distinct subset of proteins whose cellular levels are regulated by GID4 including HMGCS1, a Pro/N-degron-containing metabolic enzyme. These data reveal human GID4 Pro/N-degron targets regulated through a combination of degradative and nondegradative functions. Going forward, PFI-7 will be a valuable research tool for investigating CTLH complex biology and facilitating development of targeted protein degradation strategies that highjack CTLH E3 ligase activity.