High-throughput screening of E3 ubiquitin ligases identifies
TRIM48 as a novel negative regulator of RIG-I signaling

J 2025

High-throughput screening of E3 ubiquitin ligases identifies TRIM48 as a novel negative regulator of RIG-I signaling

WU, Guandi; Jamie FRANKISH; Joschka WILLEMSEN; Dominik RICKEN; Jonas BECKER et al.

Základní údaje

Originální název

High-throughput screening of E3 ubiquitin ligases identifies TRIM48 as a novel negative regulator of RIG-I signaling

Autoři

WU, Guandi; Jamie FRANKISH; Joschka WILLEMSEN; Dominik RICKEN; Jonas BECKER; Darius SCHWEINOCH; Sandra WUEST; Nina BEIL; Petr MATULA; Karl ROHR; Holger ERFLE; Lars KADERALI a Marco BINDER

Vydání

CELLULAR SIGNALLING, NEW YORK, ELSEVIER SCIENCE INC, 2025, 0898-6568

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10601 Cell biology

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 3.700 v roce 2024

Označené pro přenos do RIV

Ano

Organizační jednotka

Fakulta informatiky

DOI

https://doi.org/10.1016/j.cellsig.2025.111973

UT WoS

001527948800001

Klíčová slova anglicky

RIG-I signaling; Innate antiviral immunity; siRNA screening; E3 ubiquitin ligases; TRIM48

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 10. 10. 2025 13:26, doc. RNDr. Petr Matula, Ph.D.

Anotace

V originále

The retinoic acid-inducible gene-I (RIG-I) signaling is crucial for cell-intrinsic innate antiviral immunity. Upon cytosolic detection of virus-associated RNA, it triggers a cascade inducing production of potent cytokines, mainly type I and III interferons (IFNs). While effective, dysregulated responses can harm the host, requiring tight pathway control. Here, we performed a comprehensive, systematic siRNA-based high-throughput screen across 616 established and putative E3 ubiquitin ligases for their impact on RIG-I signaling. We employed a fluorescence-based live-cell imaging assay in A549 cells to monitor nuclear translocation of IRF3 and NF-kappa B, two key transcription factors downstream of RIG-I. Candidate genes were validated in an orthogonal secondary screen, assessing their impact on the functional antiviral response to a Rift Valley Fever reporter virus. Fourteen hits showed consistent effects on RIG-I signaling across both screens. These genes were further validated and characterized by assessing IFN-beta promoter reporter activity and IFNB1 mRNA levels upon dsRNA transfection. TRIM48 emerged as a highly robust negative regulator. Overexpression of TRIM48 suppressed RIG-I-mediated activation of IRF3 and NF-kappa B, reduced IFN and IFN-stimulated gene expression, and enhanced viral replication. Conversely, TRIM48 deficiency enhanced RIG-I signaling and inhibited viral replication. Notably, TRIM48 acts as an induced feedback regulator upon infection, and its effect depended on its enzymatic ubiquitin ligase activity. Our high-throughput screen provides an unbiased assessment of close to all E3 ubiquitin ligases for their regulatory effect in RIG-I signaling, and identified several interesting candidates for further investigation. TRIM48 was established as a negative feedback regulator of the RIG-I pathway.

Citovat

WU, Guandi; Jamie FRANKISH; Joschka WILLEMSEN; Dominik RICKEN; Jonas BECKER; Darius SCHWEINOCH; Sandra WUEST; Nina BEIL; Petr MATULA; Karl ROHR; Holger ERFLE; Lars KADERALI a Marco BINDER. High-throughput screening of E3 ubiquitin ligases identifies TRIM48 as a novel negative regulator of RIG-I signaling. CELLULAR SIGNALLING. NEW YORK: ELSEVIER SCIENCE INC, 2025, roč. 134, č. 1, s. 1-18. ISSN 0898-6568. Dostupné z: https://doi.org/10.1016/j.cellsig.2025.111973.

@article{2524803,
   author = {Wu, Guandi and Frankish, Jamie and Willemsen, Joschka and Ricken, Dominik and Becker, Jonas and Schweinoch, Darius and Wuest, Sandra and Beil, Nina and Matula, Petr and Rohr, Karl and Erfle, Holger and Kaderali, Lars and Binder, Marco},
   article_location = {NEW YORK},
   article_number = {1},
   doi = {https://doi.org/10.1016/j.cellsig.2025.111973},
   keywords = {RIG-I signaling; Innate antiviral immunity; siRNA screening; E3 ubiquitin ligases; TRIM48},
   language = {eng},
   issn = {0898-6568},
   journal = {CELLULAR SIGNALLING},
   title = {High-throughput screening of E3 ubiquitin ligases identifies TRIM48 as a novel negative regulator of RIG-I signaling},
   url = {https://www.sciencedirect.com/science/article/pii/S0898656825003882?via%3Dihub},
   volume = {134},
   year = {2025}
}

TY  - JOUR
ID  - 2524803
AU  - Wu, Guandi - Frankish, Jamie - Willemsen, Joschka - Ricken, Dominik - Becker, Jonas - Schweinoch, Darius - Wuest, Sandra - Beil, Nina - Matula, Petr - Rohr, Karl - Erfle, Holger - Kaderali, Lars - Binder, Marco
PY  - 2025
TI  - High-throughput screening of E3 ubiquitin ligases identifies TRIM48 as a novel negative regulator of RIG-I signaling
JF  - CELLULAR SIGNALLING
VL  - 134
IS  - 1
SP  - 1-18
EP  - 1-18
PB  - ELSEVIER SCIENCE INC
SN  - 08986568
KW  - RIG-I signaling
KW  - Innate antiviral immunity
KW  - siRNA screening
KW  - E3 ubiquitin ligases
KW  - TRIM48
UR  - https://www.sciencedirect.com/science/article/pii/S0898656825003882?via%3Dihub
N2  - The retinoic acid-inducible gene-I (RIG-I) signaling is crucial for cell-intrinsic innate antiviral immunity. Upon cytosolic detection of virus-associated RNA, it triggers a cascade inducing production of potent cytokines, mainly type I and III interferons (IFNs). While effective, dysregulated responses can harm the host, requiring tight pathway control. Here, we performed a comprehensive, systematic siRNA-based high-throughput screen across 616 established and putative E3 ubiquitin ligases for their impact on RIG-I signaling. We employed a fluorescence-based live-cell imaging assay in A549 cells to monitor nuclear translocation of IRF3 and NF-kappa B, two key transcription factors downstream of RIG-I. Candidate genes were validated in an orthogonal secondary screen, assessing their impact on the functional antiviral response to a Rift Valley Fever reporter virus. Fourteen hits showed consistent effects on RIG-I signaling across both screens. These genes were further validated and characterized by assessing IFN-beta promoter reporter activity and IFNB1 mRNA levels upon dsRNA transfection. TRIM48 emerged as a highly robust negative regulator. Overexpression of TRIM48 suppressed RIG-I-mediated activation of IRF3 and NF-kappa B, reduced IFN and IFN-stimulated gene expression, and enhanced viral replication. Conversely, TRIM48 deficiency enhanced RIG-I signaling and inhibited viral replication. Notably, TRIM48 acts as an induced feedback regulator upon infection, and its effect depended on its enzymatic ubiquitin ligase activity. Our high-throughput screen provides an unbiased assessment of close to all E3 ubiquitin ligases for their regulatory effect in RIG-I signaling, and identified several interesting candidates for further investigation. TRIM48 was established as a negative feedback regulator of the RIG-I pathway.
ER  -

WU, Guandi; Jamie FRANKISH; Joschka WILLEMSEN; Dominik RICKEN; Jonas BECKER; Darius SCHWEINOCH; Sandra WUEST; Nina BEIL; Petr MATULA; Karl ROHR; Holger ERFLE; Lars KADERALI a Marco BINDER. High-throughput screening of E3 ubiquitin ligases identifies TRIM48 as a novel negative regulator of RIG-I signaling. \textit{CELLULAR SIGNALLING}. NEW YORK: ELSEVIER SCIENCE INC, 2025, roč.~134, č.~1, s.~1-18. ISSN~0898-6568. Dostupné z: https://doi.org/10.1016/j.cellsig.2025.111973.

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