Abstract
We recently reported that hydrogen peroxide-inducible clone-5 (Hic-5, also named androgen receptor-associated protein 55) can bind to the transforming growth factor-β (TGF-β)-signaling regulator Smad3, thereby inhibiting certain Smad3-dependent TGF-β responses. We now show that Hic-5 can also control TGF-β responses through an alternative mechanism involving Smad7, a key negative regulator of TGF-β signaling. Hic-5 binds directly to Smad7. This interaction requires the LIM3 domain of Hic-5, and enhances TGF-β signaling through causing loss of Smad7 protein but not mRNA. Enforced expression of Hic-5 reverses the ability of Smad7 to suppress TGF-β-induced phosphorylation of Smads 2 and 3 and activation of the plasminogen activator inhibitor-1 promoter (in NRP-154 and PC3 prostate carcinoma and WPMY-1 prostate myofibroblast cell lines). Lentiviral-mediated small-hairpin RNA silencing of endogenous Hic-5 reduced TGF-β responses in PC3 and WPMY-1 cells. Further work suggests that the level of Smad7 is modulated by its physical interaction with Hic-5 and targeted to a degradation pathway not likely to be proteasomal. Our findings support that Hic-5 functions as a cell-type-specific activator of TGF-β signaling through its ability to physically interact with and neutralize Smad7.
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Change history
24 October 2022
A Correction to this paper has been published: https://doi.org/10.1038/s41388-022-02510-8
Abbreviations
- AR:
-
androgen receptor
- ARA55:
-
androgen receptor-associated protein 55
- ARE-lux:
-
Smad2-responsive reporter containing activin response element
- co-IP:
-
co-immunoprecipitation
- DMEM/F-12:
-
Dulbecco's modified Eagle's medium/Ham's F-12
- ECM:
-
extracellular matrix
- EMT:
-
epithelial-mesenchymal transition
- FAST-1:
-
forkhead activin signal transducer-1
- FBS:
-
fetal bovine serum
- GST:
-
glutathione S-transferase
- HEK293:
-
human embryonic kidney cell line 293
- Hic-5:
-
hydrogen peroxide-inducible clone-5
- PAI-1:
-
plasminogen activator inhibitor-1
- shRNA:
-
small-hairpin RNA
- TβRI:
-
TGF-β type I receptor
- TβRII:
-
TGF-β type II receptor
- TGF-β:
-
transforming growth factor-β
- 3TP-lux:
-
TGF-β-responsive reporter containing PAI-1 promoter region
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Acknowledgements
We thank Dr Joan Massagué for 3TP-lux plasmid, Dr Bert Vogelstein for SBE4BV-luciferase plasmid, Dr Kohei Miyazono for full-length mouse Flag-Smad7-pcDNA3 plasmid, Dr Bing-Cheng Wang for the pcDNA3-Myc plasmid, Dr Ryosuke Takahashi and Dr Yu-Chung Yang for HA-ubiquitin plasmid, Dr Malcolm Whitman for ARE-lux and Myc-FAST-1. This work was supported by NCI grants 1R01CA102074 and 1R01CA092102.
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Wang, H., Song, K., Krebs, T. et al. Smad7 is inactivated through a direct physical interaction with the LIM protein Hic-5/ARA55. Oncogene 27, 6791–6805 (2008). https://doi.org/10.1038/onc.2008.291
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DOI: https://doi.org/10.1038/onc.2008.291
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