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:

DHX15 promotes prostate cancer progression by stimulating Siah2-mediated ubiquitination of androgen receptor

Oncogene volume 37, pages 638–650 (2018)Cite this article

Subjects

Abstract

Androgen receptor (AR) activation is critical for prostate cancer (PCa) development and progression, including castration resistance. The nuclear export signal of AR (NESAR) has an important role in AR intracellular trafficking and proteasome-dependent degradation. Here, we identified the RNA helicase DHX15 as a novel AR co-activator using a yeast mutagenesis screen and revealed that DHX15 regulates AR activity by modulating E3 ligase Siah2-mediated AR ubiquitination independent of its ATPase activity. DHX15 and Siah2 form a complex with AR, through NESAR. DHX15 stabilized Siah2 and enhanced its E3 ubiquitin-ligase activity, resulting in AR activation. Importantly, DHX15 was upregulated in PCa specimens and its expression was correlated with Gleason scores and prostate-specific antigen recurrence. Furthermore, DHX15 immunostaining correlated with Siah2. Finally, DHX15 knockdown inhibited the growth of C4-2 prostate tumor xenografts in mice. Collectively, our data argue that DHX15 enhances AR transcriptional activity and contributes to PCa progression through Siah2.

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
Figure 7

Similar content being viewed by others

References

  1. Siegel RL, Miller KD, Jemal A . Cancer statistics, 2017. CA Cancer J Clin 2017; 67: 7–30.

    Article  PubMed  Google Scholar 

  2. Hellerstedt BA, Pienta KJP . The cturrent state of hormonal therapy for prostate cancer. CA Cancer J Clin 2002; 52: 154–179.

    Article  PubMed  Google Scholar 

  3. Chan SC, Dehm SM . Constitutive activity of the androgen receptor. Adv Pharmacol 2014; 70: 327–366.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Kobayashi T, Inoue T, Kamba T, Ogawa O . Experimental evidence of persistent androgen-receptor-dependency in castration-resistant prostate cancer. Int J Mol Sci 2013; 14: 15615–15635.

    Article  PubMed  PubMed Central  Google Scholar 

  5. Gelmann EP . Molecular biology of the androgen receptor. J Clin Oncol 2002; 20: 3001–3015.

    Article  CAS  PubMed  Google Scholar 

  6. Lonergan PE, Tindall DJ . Androgen receptor signaling in prostate cancer development and progression. J Carcinog 2011; 10: 20.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Picard D, Yamamoto KR . Two signals mediate hormone-dependent nuclear localization of the glucocorticoid receptor. EMBO J 1987; 6: 3333–3340.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Poukka H, Karvonen U, Yoshikawa N, Tanaka H, Palvimo JJ, Janne OA . The RING finger protein SNURF modulates nuclear trafficking of the androgen receptor. J Cell Sci 2000; 113: 2991–3001.

    CAS  PubMed  Google Scholar 

  9. Saporita AJ, Zhang Q, Navai N, Dincer Z, Hahn J, Cai X et al. Identification and characterization of a ligand-regulated nuclear export signal in androgen receptor. J Biol Chem 2003; 278: 41998–42005.

    Article  CAS  PubMed  Google Scholar 

  10. Zhou ZX, Sar M, Simental JA, Lane MV, Wilson EM . A ligand-dependent bipartite nuclear targeting signal in the human androgen receptor. Requirement for the DNA-binding domain and modulation by NH2-terminal and carboxyl-terminal sequences. J Biol Chem 1994; 269: 13115–13123.

    CAS  PubMed  Google Scholar 

  11. Gong Y, Wang D, Dar JA, Singh P, Graham L, Liu W et al. Nuclear export signal of androgen receptor (NESAR) regulation of androgen receptor level in human prostate cell lines via ubiquitination and proteasome-dependent degradation. Endocrinology 2012; 153: 5716–5725.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Nguyen MM, Harmon RM, Wang Z . Characterization of karyopherins in androgen receptor intracellular trafficking in the yeast model. Int J Clin Exp Pathol 2014; 7: 2768–2779.

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Ferraldeschi R, Welti J, Luo J, Attard G, de Bono JS . Targeting the androgen receptor pathway in castration-resistant prostate cancer: progresses and prospects. Oncogene 2015; 34: 1745–1757.

    Article  CAS  PubMed  Google Scholar 

  14. Shafi AA, Yen AE, Weigel NL . Androgen receptors in hormone-dependent and castration-resistant prostate cancer. Pharmacol Ther 2013; 140: 223–238.

    Article  CAS  PubMed  Google Scholar 

  15. Reed JC, Ely KR . Degrading liaisons: Siah structure revealed. Nat Struct Biol 2002; 9: 8–10.

    Article  CAS  PubMed  Google Scholar 

  16. Fukuba H, Takahashi T, Jin HG, Kohriyama T, Matsumoto M . Abundance of aspargynyl-hydroxylase FIH is regulated by Siah-1 under normoxic conditions. Neurosci Lett 2008; 433: 209–214.

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  18. Nadeau RJ, Toher JL, Yang X, Kovalenko D, Friesel R . Regulation of Sprouty2 stability by mammalian Seven-in-Absentia homolog 2. J Cell Biochem 2007; 100: 151–160.

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  20. Zhang J, Guenther MG, Carthew RW, Lazar MA . Proteasomal regulation of nuclear receptor corepressor-mediated repression. Genes Development 1998; 12: 1775–1780.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Nakayama K, Qi J, Ronai Z . The ubiquitin ligase Siah2 and the hypoxia response. Mol Cancer Res 2009; 7: 443–451.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Qi J, Tripathi M, Mishra R, Sahgal N, Fazli L, Ettinger S et al. The E3 ubiquitin ligase Siah2 contributes to castration-resistant prostate cancer by regulation of androgen receptor transcriptional activity. Cancer Cell 2013; 23: 332–346.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Freeman MR . The ubiquitin ligase Siah2 is revealed as an accomplice of the androgen receptor in castration resistant prostate cancer. Asian J Androl 2013; 15: 447–448.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Arenas JE, Abelson JN . Prp43: An RNA helicase-like factor involved in spliceosome disassembly. Proc Nat Acad Sci USA 1997; 94: 11798–11802.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Combs DJ, Nagel RJ, Ares M Jr, Stevens SW . Prp43p is a DEAH-box spliceosome disassembly factor essential for ribosome biogenesis. Mol Cell Biol 2006; 26: 523–534.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Tanaka N, Aronova A, Schwer B . Ntr1 activates the Prp43 helicase to trigger release of lariat-intron from the spliceosome. Genes Dev 2007; 21: 2312–2325.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Fourmann JB, Schmitzova J, Christian H, Urlaub H, Ficner R, Boon KL et al. Dissection of the factor requirements for spliceosome disassembly and the elucidation of its dissociation products using a purified splicing system. Genes Dev 2013; 27: 413–428.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Koodathingal P, Novak T, Piccirilli JA, Staley JP . The DEAH box ATPases Prp16 and Prp43 cooperate to proofread 5' splice site cleavage during pre-mRNA splicing. Mol Cell 2010; 39: 385–395.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Walbott H, Mouffok S, Capeyrou R, Lebaron S, Humbert O, van Tilbeurgh H et al. Prp43p contains a processive helicase structural architecture with a specific regulatory domain. EMBO J 2010; 29: 2194–2204.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Brown JD, Beggs JD . Roles of PRP8 protein in the assembly of splicing complexes. EMBO J 1992; 11: 3721–3729.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Mosallanejad K, Sekine Y, Ishikura-Kinoshita S, Kumagai K, Nagano T, Matsuzawa A et al. The DEAH-box RNA helicase DHX15 activates NF-kappaB and MAPK signaling downstream of MAVS during antiviral responses. Sci Signal 2014; 7: ra40.

    Article  PubMed  Google Scholar 

  32. Coffey K, Robson CN . Regulation of the androgen receptor by post-translational modifications. J Endocrinol 2012; 215: 221–237.

    Article  CAS  PubMed  Google Scholar 

  33. Chymkowitch P, Le May N, Charneau P, Compe E, Egly JM . The phosphorylation of the androgen receptor by TFIIH directs the ubiquitin/proteasome process. EMBO J 2011; 30: 468–479.

    Article  CAS  PubMed  Google Scholar 

  34. Gaughan L, Logan IR, Neal DE, Robson CN . Regulation of androgen receptor and histone deacetylase 1 by Mdm2-mediated ubiquitylation. Nucleic Acids Res 2005; 33: 13–26.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Xu K, Shimelis H, Linn DE, Jiang R, Yang X, Sun F et al. Regulation of androgen receptor transcriptional activity and specificity by RNF6-induced ubiquitination. Cancer Cell 2009; 15: 270–282.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Li B, Lu W, Chen Z . Regulation of androgen receptor by E3 ubiquitin ligases: for more or less. Receptors Clin Investig 2014; 1; doi:10.14800%2Frci.122.

  37. Hu G, Fearon ER . 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 1999; 19: 724–732.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Scortegagna M, Subtil T, Qi J, Kim H, Zhao W, Gu W et al. USP13 enzyme regulates Siah2 ligase stability and activity via noncatalytic ubiquitin-binding domains. J Biol Chem 2011; 286: 27333–27341.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Clark EL, Coulson A, Dalgliesh C, Rajan P, Nicol SM, Fleming S et al. The RNA helicase p68 is a novel androgen receptor coactivator involved in splicing and is overexpressed in prostate cancer. Cancer Res 2008; 68: 7938–7946.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Fouraux MA, Kolkman MJ, Van der Heijden A, De Jong AS, Van Venrooij WJ, Pruijn GJ . The human La (SS-B) autoantigen interacts with DDX15/hPrp43, a putative DEAH-box RNA helicase. RNA 2002; 8: 1428–1443.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Grosso AR, Martins S, Carmo-Fonseca M . The emerging role of splicing factors in cancer. EMBO Rep 2008; 9: 1087–1093.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Dong X, Sweet J, Challis JR, Brown T, Lye SJ . Transcriptional activity of androgen receptor is modulated by two RNA splicing factors, PSF and p54nrb. Mol Cell Biol 2007; 27: 4863–4875.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Rajan P, Gaughan L, Dalgliesh C, El-Sherif A, Robson CN, Leung HY et al. The RNA-binding and adaptor protein Sam68 modulates signal-dependent splicing and transcriptional activity of the androgen receptor. J Pathol 2008; 215: 67–77.

    Article  CAS  PubMed  Google Scholar 

  45. Sequoia Ecosystem and Recreation Preserve Act of 1999. 106th Congress ed, US House of Representatives1999.

  46. Haider AS, Kaye G, Thomson A . Autoimmune hepatitis in a demographically isolated area of Australia. Int MedJ 2010; 40: 281–285.

    Article  CAS  Google Scholar 

  47. Zhao HL, Ueki N, Hayman MJ . The Ski protein negatively regulates Siah2-mediated HDAC3 degradation. Biochem Biophys Res Commun 2010; 399: 623–628.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank Aiyuan Zhang for technical support, Richard Gaber (Northwestern University) for advice on yeast screen, the PCBN for TMA with PCa specimens, Megan Lambert, Krystal Roskov and Robin Frederick for animal husbandry, and members of the Wang Lab for discussion. This work was supported in part by NIH R01 CA186780 (ZW), NIH R01 CA108675 (ZW), ACS #PF-05-229-01-CSM (MMN), the Department of Defense Prostate Cancer Research Program Award No W81XWH-14-2-0182, W81XWH-14-2-0183, W81XWH-14-2-0185, W81XWH-14-2-0186, and W81XWH-15-2-0062 PCBN, National Nature Science Foundation of China #81402091 (YJ), and National Natural Science Foundation of China Key Project 81130046 (JZ) and 2013GXNSFEA053004 (JZ). This work was also supported by a post-doctoral fellowship from the Urology Care Foundation of the American Urological Association (DW), the Mellam Family Fellowship (DW and KZM), the Tippens Scholarship (LEP) and NIH R50 CA211242-01 (LEP). In addition, this research was supported by the UPCI Animal Facility, Vector Core and Tissue and Research Pathology Services (TARPS) funded through NCI CCSG P30CA047904.

Author contributions

Yifeng Jing conceived and performed the majority of experiments and wrote manuscript. Minh M Nguyen performed the yeast screening. Dan Wang constructed the plasmids of flag-DHX15 and myc-DHX15, performed radioactive ligand-binding assay and ChIP and assisted with technical details and experimental design. Laura E Pascal performed animal experiments and analysis of the animal data, and helped with manuscript. Wenhuan Guo constructed the truncated Siah2 plasmids, performed parts of BrdU assay and several extractions of proteins and RNAs. Yadong Xu generated data for Supplementary Figures S1 and S3. Junkui Ai constructed the plasmid of Flag-AR and C4-2-AR, assisted with technical details. Fang-ming Deng performed IHC. Khalid Masoodi constructed and standardized the primers of DHX15. Xinpei Yu performed some of the ubiquitination IP assays. Shujie Xia, Jian Zhang, Joel B Nelson conceived study and helped with manuscript. Zhou Wang conceived and directed study, and helped with manuscript.

Author information

Authors and Affiliations

  1. Department of Urology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, PR China

    Y Jing & S Xia

  2. Department of Urology, University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA,

    Y Jing, M M Nguyen, D Wang, L E Pascal, W Guo, Y Xu, J Ai, K Z Masoodi, J B Nelson & Z Wang

  3. Department of Pathology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, PR China

    W Guo

  4. Department of Urology, The Second Xiangya Hospital of Central South University, Hunan, China

    Y Xu

  5. The third Xiangya Hospital of Central South University, Changsha, China

    Y Xu

  6. Department of Pathology, NYU School of Medicine, New York, NY, USA

    F-M Deng

  7. Division of Plant Biotechnology, Transcriptomics Lab, SKUAST-K, Shalimar, Srinagar, J&K, India

    K Z Masoodi

  8. Department of Geriatrics, Guangzhou General Hospital of Guangzhou Military Command,

    X Yu

  9. Guangdong Provincial Key Laboratory of Geriatric Infection and Organ Function Support, Guangzhou, Guangdong, China

    X Yu

  10. Cancer Center, Traditional Chinese Medicine-Integrated Hospital, Southern Medical University, Guangzhou, Guangdong, China

    X Yu

  11. Center for Translational Medicine, Guangxi Medical University, Nanning, Guangxi, China

    J Zhang

  12. University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    J Zhang

  13. Department of Molecular Pharmacology and Chemical Biology, University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA,

    J B Nelson & Z Wang

  14. Department of Pathology, University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA,

    Z Wang

Authors
  1. Y Jing
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M M Nguyen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. D Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. L E Pascal
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. W Guo
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Y Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. J Ai
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. F-M Deng
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. K Z Masoodi
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. X Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. J Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. J B Nelson
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. S Xia
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Z Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to S Xia or Z Wang.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies this paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Jing, Y., Nguyen, M., Wang, D. et al. DHX15 promotes prostate cancer progression by stimulating Siah2-mediated ubiquitination of androgen receptor. Oncogene 37, 638–650 (2018). https://doi.org/10.1038/onc.2017.371

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2017.371

This article is cited by

Search

Advanced search

Quick links