Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Siah-1S, a novel splice variant of Siah-1 (seven in absentia homolog), counteracts Siah-1-mediated downregulation of β-catenin

Oncogene volume 26, pages 6319–6331 (2007)Cite this article

Abstract

Siah-1 (seven in absentia homolog) is known to cause indirect degradation of β-catenin through formation of a complex with Siah-interacting protein (SIP), Skp1 and Ebi. Here, we report the characterization of a novel splice variant of human Siah-1, designated Siah-1S, which is produced by an alternative splicing mechanism. The novel intron/exon junctions used to generate Siah-1S follow a non-conventional CT-AC rule. Siah-1S exhibits an even shorter half-life than Siah-1 and is able to catalyse self-ubiquitination that results in its subsequent degradation by proteasome. Siah-1S is shown to upregulate β-catenin-dependent Tcf/Lef transcriptional activation and antagonize Siah-1's potentiation effect on the apoptosis induced by etoposide in MCF-7 cells. Additionally, Siah-1S is found to interact with Siah-1 to form heterodimer or with itself to form homodimer. Unlike homodimer Siah-1*Siah-1, neither Siah-1*Siah-1S nor Siah-1S*Siah-1S is able to bind to Siah-1-interacting protein, which may explain the underlying mechanism for Siah-1S's dominant negative effect on Siah-1. Importantly, results from in vitro soft agar assay demonstrated that Siah-1S displays a promotion effect on cells tumorigenicity.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

Accession codes

Accessions

GenBank/EMBL/DDBJ

References

  • Amson RB, Nemani M, Roperch JP, Israeli D, Bougueleret L, Le Gall I et al. (1996). Isolation of 10 differentially expressed cDNAs in p53-induced apoptosis: activation of the vertebrate homologue of the drosophila seven in absentia gene. Proc Natl Acad Sci USA 93: 3953–3957.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bienz M, Clevers H . (2000). Linking colorectal cancer to Wnt signaling. Cell 103: 311–320.

    Article  CAS  PubMed  Google Scholar 

  • Boehm J, He Y, Greiner A, Staudt L, Wirth T . (2001). Regulation of BOB.1/OBF.1 stability by SIAH. EMBO J 20: 4153–4162.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bruzzoni-Giovanelli H, Faille A, Linares-Cruz G, Nemani M, Le Deist F, Germani A et al. (1999). SIAH-1 inhibits cell growth by altering the mitotic process. Oncogene 18: 7101–7109.

    Article  CAS  PubMed  Google Scholar 

  • Carthew RW, Rubin GM . (1990). Seven in absentia, a gene required for specification of R7 cell fate in the Drosophila eye. Cell 63: 561–577.

    Article  CAS  PubMed  Google Scholar 

  • Dong X, Tsuda L, Zavitz kH, Lin M, Li S, Carthew RW et al. (1999). ebi regulates epidermal growth factor receptor signaling pathways in Drosophila. Genes Dev 13: 954–965.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • French C, Menegazzi P, Nicholson L, Macaulay H, DiLuca D, Gompels UA . (1999). Novel, nonconsensus cellular splicing regulates expression of a gene encoding a chemokine-like protein that shows high variation and is specific for human herpesvirus 6. Virology 262: 139–151.

    Article  CAS  PubMed  Google Scholar 

  • Fukushima T, Zapata JM, Singha NC, Thomas M, Kress CL, Krajewska M et al. (2006). Critical function for SIP, a ubiquitin E3 ligase component of the beta-catenin degradation pathway, for thymocyte development and G1 checkpoint. Immunity 24: 29–39.

    Article  CAS  PubMed  Google Scholar 

  • Germani A, Bruzzoni-Giovanelli H, Fellous A, Gisselbrecht S, Varin-Blank N, Calvo F . (2000). SIAH-1 interacts with alpha-tubulin and degrades the kinesin Kid by the proteasome pathway during mitosis. Oncogene 19: 5997–6006.

    Article  CAS  PubMed  Google Scholar 

  • Germani A, Prabel A, Mourah S, Podgorniak MP, Di Carlo A, Ehrlich R et al. (2003). SIAH-1 interacts with CtIP and promotes its degradation by the proteasome pathway. Oncogene 22: 8845–8851.

    Article  CAS  PubMed  Google Scholar 

  • Germani A, Romero F, Houlard M, Camonis J, Gisselbrecht S, Fischer S et al. (1999). hSiah2 is a new Vav binding protein which inhibits Vav-mediated signaling pathways. Mol Cell Biol 19: 3798–3807.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gutierrez GJ, Vogtlin A, Castro A, Ferby I, Salvagiotto G, Ronai Z et al. (2006). Meiotic regulation of the CDK activator RINGO/Speedy by ubiquitin-proteasome-mediated processing and degradation. Nat Cell Biol 8: 1084–1094.

    Article  CAS  PubMed  Google Scholar 

  • Habelhah H, Laine A, Erdjument-Bromage H, Tempst P, Gershwin ME, Bowtell DD et al. (2004). Regulation of 2-oxoglutarate (alpha-ketoglutarate) dehydrogenase stability by the RING finger ubiquitin ligase Siah. J Biol Chem 279: 53782–53788.

    Article  CAS  PubMed  Google Scholar 

  • Haupt Y, Maya R, Kazaz A, Oren M . (1997). Mdm2 promotes the rapid degradation of p53. Nature 387: 296–299.

    Article  CAS  PubMed  Google Scholar 

  • He TC, Sparks AB, Rago C, Hermeking H, Zawel L, da Costa LT et al. (1998). Identification of c-MYC as a target of the APC pathway. Science 281: 1509–1512.

    Article  CAS  PubMed  Google Scholar 

  • Hu G, Chung YL, Glover T, Valentine V, Look AT, Fearon ER . (1997a). Characterization of human homologs of the Drosophila seven in absentia (sina) gene. Genomics 46: 103–111.

    Article  CAS  PubMed  Google Scholar 

  • Hu G, Fearon ER . (1999). Siah-1 N-terminal RING domain is required for proteolysis function, and C-terminal sequences regulate oligomerization and binding to target proteins. Mol Cell Biol 19: 724–732.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hu G, Zhang S, Vidal M, Baer JL, Xu T, Fearon ER . (1997b). Mammalian homologs of seven in absentia regulate DCC via the ubiquitin–proteasome pathway. Genes Dev 11: 2701–2714.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Iwai A, Marusawa H, Matsuzawa S, Fukushima T, Hijikata M, Reed JC et al. (2004). Siah-1L, a novel transcript variant belonging to the human Siah family of proteins, regulates beta-catenin activity in a p53-dependent manner. Oncogene 23: 7593–7600.

    Article  CAS  PubMed  Google Scholar 

  • Johnsen SA, Subramaniam M, Monroe DG, Janknecht R, Spelsberg TC . (2002). Modulation of transforming growth factor beta (TGFbeta)/Smad transcriptional responses through targeted degradation of TGFbeta-inducible early gene-1 by human seven in absentia homologue. J Biol Chem 277: 30754–30759.

    Article  CAS  PubMed  Google Scholar 

  • Kim H, Jeong W, Ahn K, Ahn C, Kang S . (2004). Siah-1 interacts with the intracellular region of polycystin-1 and affects its stability via the ubiquitin–proteasome pathway. J Am Soc Nephrol 15: 2042–2049.

    Article  CAS  PubMed  Google Scholar 

  • Kramer A . (1996). The structure and function of proteins involved in mammalian pre-mRNA splicing. Annu Rev Biochem 65: 367–409.

    Article  CAS  PubMed  Google Scholar 

  • Kubbutat MH, Jones SN, Vousden KH . (1997). Regulation of p53 stability by Mdm2. Nature 387: 299–303.

    Article  CAS  PubMed  Google Scholar 

  • Li S, Li Y, Carthew RW, Lai ZC . (1997). Photoreceptor cell differentiation requires regulated proteolysis of the transcriptional repressor Tramtrack. Cell 90: 469–478.

    Article  CAS  PubMed  Google Scholar 

  • Liu J, Stevens J, Rote CA, Yost HJ, Hu Y, Neufeld KL et al. (2001). Siah-1 mediates a novel beta-catenin degradation pathway linking p53 to the adenomatous polyposis coli protein. Mol Cell 7: 927–936.

    Article  CAS  PubMed  Google Scholar 

  • Lorick KL, Jensen JP, Fang S, Ong AM, Hatakeyama S, Weissman AM . (1999). RING fingers mediate ubiquitin-conjugating enzyme (E2)-dependent ubiquitination. Proc Natl Acad Sci USA 96: 11364–11369.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Matsuzawa S, Li C, Ni CZ, Takayama S, Reed JC, Ely KR . (2003). Structural analysis of Siah1 and its interactions with Siah-interacting protein (SIP). J Biol Chem 278: 1837–1840.

    Article  CAS  PubMed  Google Scholar 

  • Matsuzawa S, Takayama S, Froesch BA, Zapata JM, Reed JC . (1998). p53-inducible human homologue of Drosophila seven in absentia (Siah) inhibits cell growth: suppression by BAG-1. EMBO J 17: 2736–2747.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Matsuzawa SI, Reed JC . (2001). Siah-1, SIP, and Ebi collaborate in a novel pathway for beta-catenin degradation linked to p53 responses. Mol Cell 7: 915–926.

    Article  CAS  PubMed  Google Scholar 

  • Mei Y, Du W, Yang Y, Wu M . (2005). Puma(*)Mcl-1 interaction is not sufficient to prevent rapid degradation of Mcl-1. Oncogene 24: 7224–7237.

    Article  CAS  PubMed  Google Scholar 

  • Nagano Y, Yamashita H, Takahashi T, Kishida S, Nakamura T, Iseki E et al. (2003). Siah-1 facilitates ubiquitination and degradation of synphilin-1. J Biol Chem 278: 51504–51514.

    Article  CAS  PubMed  Google Scholar 

  • Nakayama K, Frew IJ, Hagensen M, Skals M, Habelhah H, Bhoumik A et al. (2004). Siah2 regulates stability of prolyl-hydroxylases, controls HIF1alpha abundance, and modulates physiological responses to hypoxia. Cell 117: 941–952.

    Article  CAS  PubMed  Google Scholar 

  • Nemani M, Linares-Cruz G, Bruzzoni-Giovanelli H, Roperch JP, Tuynder M, Bougueleret L et al. (1996). Activation of the human homologue of the Drosophila sina gene in apoptosis and tumor suppression. Proc Natl Acad Sci USA 93: 9039–9042.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Oliver PL, Bitoun E, Clark J, Jones EL, Davies KE . (2004). Mediation of Af4 protein function in the cerebellum by Siah proteins. Proc Natl Acad Sci USA 101: 14901–14906.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Peifer M, Polakis P . (2000). Wnt signaling in oncogenesis and embryogenesis — a look outside the nucleus. Science 287: 1606–1609.

    Article  CAS  PubMed  Google Scholar 

  • Polekhina G, House CM, Traficante N, Mackay JP, Relaix F, Sassoon DA et al. (2002). Siah ubiquitin ligase is structurally related to TRAF and modulates TNF-alpha signaling. Nat Struct Biol 9: 68–75.

    Article  CAS  PubMed  Google Scholar 

  • Provost E, Rimm DL . (1999). Controversies at the cytoplasmic face of the cadherin-based adhesion complex. Curr Opin Cell Biol 11: 567–572.

    Article  CAS  PubMed  Google Scholar 

  • Roperch JP, Lethrone F, Prieur S, Piouffre L, Israeli D, Tuynder M et al. (1999). SIAH-1 promotes apoptosis and tumor suppression through a network involving the regulation of protein folding, unfolding, and trafficking: identification of common effectors with p53 and p21(Waf1). Proc Natl Acad Sci USA 96: 8070–8073.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Santelli E, Leone M, Li C, Fukushima T, Preece NE, Olson AJ et al. (2005). Structural analysis of Siah1-Siah-interacting protein interactions and insights into the assembly of an E3 ligase multiprotein complex. J Biol Chem 280: 34278–34287.

    Article  CAS  PubMed  Google Scholar 

  • Susini L, Passer BJ, Amzallag-Elbaz N, Juven-Gershon T, Prieur S, Privat N et al. (2001). Siah-1 binds and regulates the function of Numb. Proc Natl Acad Sci USA 98: 15067–15072.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tang AH, Neufeld TP, Kwan E, Rubin GM . (1997). PHYL acts to downregulate TTK88, a transcriptional repressor of neuronal cell fates, by a SINA-dependent mechanism. Cell 90: 459–467.

    Article  CAS  PubMed  Google Scholar 

  • Tanikawa J, Ichikawa-Iwata E, Kanei-Ishii C, Nakai A, Matsuzawa S, Reed JC et al. (2000). p53 suppresses the c-Myb-induced activation of heat shock transcription factor 3. J Biol Chem 275: 15578–15585.

    Article  CAS  PubMed  Google Scholar 

  • Tetsu O, McCormick F . (1999). Beta-catenin regulates expression of cyclin D1 in colon carcinoma cells. Nature 398: 422–426.

    Article  CAS  PubMed  Google Scholar 

  • Tiedt R, Bartholdy BA, Matthias G, Newell JW, Matthias P . (2001). The RING finger protein Siah-1 regulates the level of the transcriptional co-activator OBF-1. EMBO J 20: 4143–4152.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tuynder M, Susini L, Prieur S, Besse S, Fiucci G, Amson R et al. (2002). Biological models and genes of tumor reversion: cellular reprogramming through tpt1/TCTP and SIAH-1. Proc Natl Acad Sci USA 99: 14976–14981.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Venables JP, Dalgliesh C, Paronetto MP, Skitt L, Thornton JK, Saunders PT et al. (2004). SIAH1 targets the alternative splicing factor T-STAR for degradation by the proteasome. Hum Mol Genet 13: 1525–1534.

    Article  CAS  PubMed  Google Scholar 

  • Wheeler TC, Chin LS, Li Y, Roudabush FL, Li L . (2002). Regulation of synaptophysin degradation by mammalian homologues of seven in absentia. J Biol Chem 277: 10273–10282.

    Article  CAS  PubMed  Google Scholar 

  • Yang Y, Fang S, Jensen JP, Weissman AM, Ashwell JD . (2000). Ubiquitin protein ligase activity of IAPs and their degradation in proteasomes in response to apoptotic stimuli. Science 288: 874–877.

    Article  CAS  PubMed  Google Scholar 

  • Zhang J, Guenther MG, Carthew RW, Lazar MA . (1998). Proteasomal regulation of nuclear receptor corepressor-mediated repression. Genes Dev 12: 1775–1780.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We are grateful to Dr Shu-ichi Matsuzawa for pTOP-FLASH, pFOP-FLASH plasmids and constructive suggestions. We also thank Dr John C Reed for providing β-catenin cDNA, and Dr Jin Lei for his technical help. This research was supported by grants from the National Natural Science Foundation of China (30530200 and 30121001), grants from the Ministry of Science and Technology of China (2002CB713702 and 2006CB910300) and a grant from Chinese Academy of Sciences (KSCX1-YW-R-57).

Author information

Authors and Affiliations

  1. Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Hefei, Anhui, People's Republic of China

    Y Mei, C Xie, W Xie & M Wu

  2. Department of Pathology, Anhui Medical University, Hefei, Anhui, People's Republic of China

    Z Wu

Authors
  1. Y Mei
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C Xie
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. W Xie
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Z Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to M Wu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mei, Y., Xie, C., Xie, W. et al. Siah-1S, a novel splice variant of Siah-1 (seven in absentia homolog), counteracts Siah-1-mediated downregulation of β-catenin. Oncogene 26, 6319–6331 (2007). https://doi.org/10.1038/sj.onc.1210449

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1210449

Keywords

This article is cited by

Search

Advanced search

Quick links