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bioRxiv · 10.1101/2023.03.23.533924

Human USP18 protects diverse cancer lineages from Type I Interferon independently of its canonical catalytic function

Abstract

Precise temporal regulation of Type I interferon signaling is imperative to effectively fight infections and cancerous cells without triggering autoimmunity. The key negative regulator of Type I interferon signaling is ubiquitin-specific protease 18 (USP18). USP18 cleaves interferon-inducible ubiquitin-like modifications through its canonical catalytic function and directly inhibits interferon receptor signaling through its scaffold role. USP18 loss-of-function dramatically impacts autoimmune disease, viral susceptibility, and cancer cell survival. However, the relative contribution of catalytic versus scaffold function is unresolved and must be determined to design effective therapeutics targeting USP18. To precisely delineate individual contribution, we evaluated the functional impact of single amino acid mutations that disrupt catalytic or scaffold activity. Here we demonstrate catalytic activity does not contribute to cell autonomous Type I interferon sensitivity across multiple cancer cell lineages. Furthermore, introducing a patient-derived mutation that disrupts scaffold function is sufficient to inhibit cancer growth. These findings establish a fundamental mechanistic basis for USP18 therapeutic design across diseases. OVERVIEWO_LIUSP18 is the key negative regulator of Type I interferon signaling in humans, mediating autoimmune disease, viral susceptibility, and cancer cell survival. C_LIO_LIUSP18 cleaves interferon-inducible ubiquitin-like modifications through its canonical catalytic function and attenuates interferon receptor signaling through its scaffold role. C_LIO_LIDelineating the contribution of each function is critical to resolve the mechanistic basis of interferon regulation and enable the development of therapeutics targeting USP18. C_LIO_LIWe demonstrate that cell intrinsic interferon sensitivity is not mediated by loss of catalytic activity. However, disruption of scaffold function by a patient-specific mutation inhibits cancer cell growth. C_LIO_LIFurthermore, we discovered that canonical catalytic activity is surprisingly inefficient in human cells. C_LIO_LIThese results clarify a fundamental mechanism of immune regulation and cancer cell survival in humans. C_LI O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=117 SRC="FIGDIR/small/533924v3_ufig1.gif" ALT="Figure 1"> View larger version (26K): org.highwire.dtl.DTLVardef@d0a353org.highwire.dtl.DTLVardef@192a09dorg.highwire.dtl.DTLVardef@16eb296org.highwire.dtl.DTLVardef@8e98a3_HPS_FORMAT_FIGEXP M_FIG C_FIG

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BibTeXRIS

Wes, P. D., Kessler, B. M., Pinto-Fernandez, A., Jove, V., Wheeler, H., Lee, C. W., Healy, D. R., Levine, K., Ralph, E. C., Giddabasappa, A., Loria, P., Yamaguchi, M., Casimiro-Garcia, A., Frattini, V., Wang, F., Yang, B.. 2023-03-25. Human USP18 protects diverse cancer lineages from Type I Interferon independently of its canonical catalytic function. https://doi.org/10.1101/2023.03.23.533924

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