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Molecular Cancer
Published in: Molecular Cancer 1/2011

Open Access 01-12-2011 | Research

Possible role of death receptor-mediated apoptosis by the E3 ubiquitin ligases Siah2 and POSH

Authors: Perry A Christian, Michael V Fiandalo, Steven R Schwarze

Published in: Molecular Cancer | Issue 1/2011

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Abstract

Background

A functioning ubiquitin proteasome system (UPS) is essential for a number of diverse cellular processes and maintenance of overall cellular homeostasis. The ability of proteasome inhibitors, such as Velcade, to promote extrinsic apoptotic effects illustrates the importance of the ubiquitin proteasome system in the regulation of death receptor signaling. Here, we set out to define the UPS machinery, particularly the E3 ubiquitin ligases, that repress apoptosis through the extrinsic pathway. A cell-based genome-wide E3 ligase siRNA screen was established to monitor caspase-8 activity following the addition of TRAIL.

Results

Data from the high-throughput screen revealed that targeting the RING-finger containing E3 ligase Siah2 as well as the signaling platform molecule POSH (SH3RF1) conferred robust caspase-8 activation in response to TRAIL stimulus. Silencing Siah2 or POSH in prostate cancer cells led to increased caspase activity and apoptosis in response to both TRAIL and Fas ligand. The E3 activity of Siah2 was responsible for mediating apoptosis resistance; while POSH protein levels were critical for maintaining viability. Further characterization of Siah2 revealed it to function downstream of early death receptor events in the apoptotic pathway. The observed apoptosis resistance provides one biological explanation for the induction of Siah2 and POSH reported in lung and prostate cancer, respectively. Expanding on an initial yeast-two-hybrid screen we have confirmed a physical interaction between E3 ligases Siah2 and POSH. Utilizing a yeast-two-hybrid mapping approach we have defined the spacer region of POSH, more specifically the RPxAxVxP motif encompassing amino acids 601-607, to be the site of Siah2 binding.

Conclusions

The data presented here define POSH and Siah2 as important mediators of death receptor mediated apoptosis and suggest targeting the interaction of these two E3 ligases is a promising novel cancer therapeutic strategy.
Appendix
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Literature
1.
Thompson CB: Apoptosis in the pathogenesis and treatment of disease. Science. 1995, 267: 1456-1462. 10.1126/science.7878464CrossRefPubMed
2.
Mitsiades N, Mitsiades CS, Richardson PG, Poulaki V, Tai YT, Chauhan D, Fanourakis G, Gu X, Bailey C, Joseph M: The proteasome inhibitor PS-341 potentiates sensitivity of multiple myeloma cells to conventional chemotherapeutic agents: therapeutic applications. Blood. 2003, 101: 2377-2380. 10.1182/blood-2002-06-1768CrossRefPubMed
3.
Vassilev LT, Vu BT, Graves B, Carvajal D, Podlaski F, Filipovic Z, Kong N, Kammlott U, Lukacs C, Klein C: In vivo activation of the p53 pathway by small-molecule antagonists of MDM2. Science. 2004, 303: 844-848. 10.1126/science.1092472CrossRefPubMed
4.
Yang Y, Yu X: Regulation of apoptosis: the ubiquitous way. Faseb J. 2003, 17: 790-799. 10.1096/fj.02-0654revCrossRefPubMed
5.
Shiozaki EN, Shi Y: Caspases, IAPs and Smac/DIABLO: mechanisms from structural biology. Trends Biochem Sci. 2004, 29: 486-494. 10.1016/j.tibs.2004.07.003CrossRefPubMed
6.
Fulda S, Wick W, Weller M, Debatin KM: Smac agonists sensitize for Apo2L/TRAIL- or anticancer drug-induced apoptosis and induce regression of malignant glioma in vivo. Nat Med. 2002, 8: 808-815.PubMed
7.
Ashkenazi A: Targeting death and decoy receptors of the tumour-necrosis factor superfamily. Nat Rev Cancer. 2002, 2: 420-430. 10.1038/nrc821CrossRefPubMed
8.
Grassme H, Cremesti A, Kolesnick R, Gulbins E: Ceramide-mediated clustering is required for CD95-DISC formation. Oncogene. 2003, 22: 5457-5470. 10.1038/sj.onc.1206540CrossRefPubMed
9.
Medema JP, Scaffidi C, Kischkel FC, Shevchenko A, Mann M, Krammer PH, Peter ME: FLICE is activated by association with the CD95 death-inducing signaling complex (DISC). Embo J. 1997, 16: 2794-2804. 10.1093/emboj/16.10.2794PubMedCentralCrossRefPubMed
10.
Thorpe JA, Christian PA, Schwarze SR: Proteasome inhibition blocks caspase-8 degradation and sensitizes prostate cancer cells to death receptor-mediated apoptosis. Prostate. 2008, 68: 200-209. 10.1002/pros.20706CrossRefPubMed
11.
Christian PA, Thorpe JA, Schwarze SR: Velcade sensitizes prostate cancer cells to TRAIL induced apoptosis and suppresses tumor growth in vivo. Cancer Biol Ther. 2009, 8: 73-80.CrossRefPubMed
12.
Borden KL: RING domains: master builders of molecular scaffolds?. J Mol Biol. 2000, 295: 1103-1112. 10.1006/jmbi.1999.3429CrossRefPubMed
13.
Xu Z, Sproul A, Wang W, Kukekov N, Greene LA: Siah1 interacts with the scaffold protein POSH to promote JNK activation and apoptosis. J Biol Chem. 2006, 281: 303-312.CrossRefPubMed
14.
Reed JC, Ely KR: Degrading liaisons: Siah structure revealed. Nat Struct Biol. 2002, 9: 8-10. 10.1038/nsb0102-8CrossRefPubMed
15.
Nakayama K, Frew IJ, Hagensen M, Skals M, Habelhah H, Bhoumik A, Kadoya T, Erdjument-Bromage H, Tempst P, Frappell PB: Siah2 regulates stability of prolyl-hydroxylases, controls HIF1alpha abundance, and modulates physiological responses to hypoxia. Cell. 2004, 117: 941-952. 10.1016/j.cell.2004.06.001CrossRefPubMed
16.
Calzado MA, de la Vega L, Moller A, Bowtell DD, Schmitz ML: An inducible autoregulatory loop between HIPK2 and Siah2 at the apex of the hypoxic response. Nat Cell Biol. 2009, 11: 85-91. 10.1038/ncb1816CrossRefPubMed
17.
Ahmed AU, Schmidt RL, Park CH, Reed NR, Hesse SE, Thomas CF, Molina JR, Deschamps C, Yang P, Aubry MC, Tang AH: Effect of disrupting seven-in-absentia homolog 2 function on lung cancer cell growth. J Natl Cancer Inst. 2008, 100: 1606-1629. 10.1093/jnci/djn365PubMedCentralCrossRefPubMed
18.
Kim GH, Park E, Kong YY, Han JK: Novel function of POSH, a JNK scaffold, as an E3 ubiquitin ligase for the Hrs stability on early endosomes. Cell Signal. 2006, 18: 553-563. 10.1016/j.cellsig.2005.05.026CrossRefPubMed
19.
Xu Z, Kukekov NV, Greene LA: POSH acts as a scaffold for a multiprotein complex that mediates JNK activation in apoptosis. EMBO J. 2003, 22: 252-261. 10.1093/emboj/cdg021PubMedCentralCrossRefPubMed
20.
Tapon N, Nagata K, Lamarche N, Hall A: A new rac target POSH is an SH3-containing scaffold protein involved in the JNK and NF-kappaB signalling pathways. EMBO J. 1998, 17: 1395-1404. 10.1093/emboj/17.5.1395PubMedCentralCrossRefPubMed
21.
Habelhah H, Frew IJ, Laine A, Janes PW, Relaix F, Sassoon D, Bowtell DD, Ronai Z: Stress-induced decrease in TRAF2 stability is mediated by Siah2. EMBO J. 2002, 21: 5756-5765. 10.1093/emboj/cdf576PubMedCentralCrossRefPubMed
22.
Hofmann TG, Moller A, Sirma H, Zentgraf H, Taya Y, Droge W, Will H, Schmitz ML: Regulation of p53 activity by its interaction with homeodomain-interacting protein kinase-2. Nat Cell Biol. 2002, 4: 1-10. 10.1038/ncb715CrossRefPubMed
23.
D'Orazi G, Cecchinelli B, Bruno T, Manni I, Higashimoto Y, Saito S, Gostissa M, Coen S, Marchetti A, Del Sal G: Homeodomain-interacting protein kinase-2 phosphorylates p53 at Ser 46 and mediates apoptosis. Nat Cell Biol. 2002, 4: 11-19. 10.1038/ncb714CrossRefPubMed
24.
Hu G, Chung YL, Glover T, Valentine V, Look AT, Fearon ER: Characterization of human homologs of the Drosophila seven in absentia (sina) gene. Genomics. 1997, 46: 103-111. 10.1006/geno.1997.4997CrossRefPubMed
25.
Schmidt M, Hanna J, Elsasser S, Finley D: Proteasome-associated proteins: regulation of a proteolytic machine. Biol Chem. 2005, 386: 725-737. 10.1515/BC.2005.085CrossRefPubMed
26.
Andersson A, Ritz C, Lindgren D, Eden P, Lassen C, Heldrup J, Olofsson T, Rade J, Fontes M, Porwit-Macdonald A: Microarray-based classification of a consecutive series of 121 childhood acute leukemias: prediction of leukemic and genetic subtype as well as of minimal residual disease status. Leukemia. 2007, 21: 1198-1203. 10.1038/sj.leu.2404688CrossRefPubMed
27.
Bhattacharjee A, Richards WG, Staunton J, Li C, Monti S, Vasa P, Ladd C, Beheshti J, Bueno R, Gillette M: Classification of human lung carcinomas by mRNA expression profiling reveals distinct adenocarcinoma subclasses. Proc Natl Acad Sci USA. 2001, 98: 13790-13795. 10.1073/pnas.191502998PubMedCentralCrossRefPubMed
28.
Lapointe J, Li C, Higgins JP, van de Rijn M, Bair E, Montgomery K, Ferrari M, Egevad L, Rayford W, Bergerheim U: Gene expression profiling identifies clinically relevant subtypes of prostate cancer. Proc Natl Acad Sci USA. 2004, 101: 811-816. 10.1073/pnas.0304146101PubMedCentralCrossRefPubMed
29.
Vanaja DK, Cheville JC, Iturria SJ, Young CY: Transcriptional silencing of zinc finger protein 185 identified by expression profiling is associated with prostate cancer progression. Cancer Res. 2003, 63: 3877-3882.PubMed
30.
Tomlins SA, Mehra R, Rhodes DR, Cao X, Wang L, Dhanasekaran SM, Kalyana-Sundaram S, Wei JT, Rubin MA, Pienta KJ: Integrative molecular concept modeling of prostate cancer progression. Nat Genet. 2007, 39: 41-51. 10.1038/ng1935CrossRefPubMed
31.
Keane MM, Ettenberg SA, Nau MM, Russell EK, Lipkowitz S: Chemotherapy augments TRAIL-induced apoptosis in breast cell lines. Cancer Res. 1999, 59: 734-741.PubMed
32.
Schwarze SR, Fu VX, Jarrard DF: Cdc37 enhances proliferation and is necessary for normal human prostate epithelial cell survival. Cancer Res. 2003, 63: 4614-4619.PubMed
33.
Peterson FC, Thorpe JA, Harder AG, Volkman BF, Schwarze SR: Structural determinants involved in the regulation of CXCL14/BRAK expression by the 26 S proteasome. J Mol Biol. 2006, 363: 813-822. 10.1016/j.jmb.2006.08.057PubMedCentralCrossRefPubMed
34.
Swift S, Lorens J, Achacoso P, Nolan GP: Rapid production of retroviruses for efficient gene delivery to mammalian cells using 293T cell-based systems. Curr Protoc Immunol. 2001, Chapter 10 (Unit 10): 17C-PubMed
Metadata
Title
Possible role of death receptor-mediated apoptosis by the E3 ubiquitin ligases Siah2 and POSH
Authors
Perry A Christian
Michael V Fiandalo
Steven R Schwarze
Publication date
01-12-2011
Publisher
BioMed Central
Published in
Molecular Cancer / Issue 1/2011
Electronic ISSN: 1476-4598
DOI
https://doi.org/10.1186/1476-4598-10-57

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