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:

Signal peptide peptidase-mediated nuclear localization of heme oxygenase-1 promotes cancer cell proliferation and invasion independent of its enzymatic activity

Oncogene volume 34, pages 2360–2370 (2015)Cite this article

Subjects

A Corrigendum to this article was published on 30 April 2015

This article has been updated

Abstract

Heme oxygenase-1 (HO-1) is a heme-degrading enzyme anchored in the endoplasmic reticulum by a carboxyl-terminal transmembrane segment (TMS). HO-1 is highly expressed in various cancers and its nuclear localization is associated with the progression of some cancers. Nevertheless, the mechanism underlying HO-1 nuclear translocation and its pathological significance remain elusive. Here we show that the signal peptide peptidase (SPP) catalyzes the intramembrane cleavage of HO-1. Coexpression of HO-1 with wild-type SPP, but not a dominant-negative SPP, promoted the nuclear localization of HO-1 in cells. Mass spectrometry analysis of cytosolic HO-1 isolated from HeLa cells overexpressing HO-1 and SPP revealed two adjacent intramembrane cleavage sites located after S275 and F276 within the TMS. Mutations of S275F276 to A275L276 significantly hindered SPP-mediated HO-1 cleavage and nuclear localization. Nuclear HO-1 was detected in A549 and DU145 cancer cell lines expressing high levels of endogenous HO-1 and SPP. SPP knockdown or inhibition significantly reduced nuclear HO-1 localization in A549 and DU145 cells. The positive nuclear HO-1 stain was also evident in lung cancer tissues expressing high levels of HO-1 and SPP. Overexpression of a truncated HO-1 (t-HO-1) lacking the TMS in HeLa and H1299 cells promoted cell proliferation and migration/invasion. The effect of t-HO-1 was not affected by a mutation in the catalytic site. However, blockade of t-HO-1 nuclear localization abolished t-HO-1-mediated effect. The tumorigenic effect of t-HO-1 was also demonstrated in the mouse model. These findings disclose that SPP-mediated intramembrane cleavage of HO-1 promotes HO-1 nuclear localization and cancer progression independent of HO-1 enzymatic activity.

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
Figure 8

Similar content being viewed by others

Change history

  • 30 April 2015

    This article has been corrected since Advance Online Publication and a corrigendum is also printed in this issue

References

  1. Abraham NG, Kappas A . Pharmacological and clinical aspects of heme oxygenase. Pharmacol Rev 2008; 60: 79–127.

    Article  CAS  Google Scholar 

  2. Jozkowicz A, Was H, Dulak J . Heme oxygenase-1 in tumors: is it a false friend? Antioxid Redox Signal 2007; 9: 2099–2117.

    Article  CAS  Google Scholar 

  3. Sunamura M, Duda DG, Ghattas MH, Lozonschi L, Motoi F, Yamauchi J et al. Heme oxygenase-1 accelerates tumor angiogenesis of human pancreatic cancer. Angiogenesis 2003; 6: 15–24.

    Article  CAS  Google Scholar 

  4. Mayerhofer M, Florian S, Krauth MT, Aichberger KJ, Bilban M, Marculescu R et al. Identification of heme oxygenase-1 as a novel BCR/ABL-dependent survival factor in chronic myeloid leukemia. Cancer Res 2004; 64: 3148–3154.

    Article  CAS  Google Scholar 

  5. Was H, Cichon T, Smolarczyk R, Rudnicka D, Stopa M, Chevalier C et al. Overexpression of heme oxygenase-1 in murine melanoma: increased proliferation and viability of tumor cells, decreased survival of mice. Am J Pathol 2006; 169: 2181–2198.

    Article  CAS  Google Scholar 

  6. Liu ZM, Chen GG, Ng EK, Leung WK, Sung JJ, Chung SC . Upregulation of heme oxygenase-1 and p21 confers resistance to apoptosis in human gastric cancer cells. Oncogene 2004; 23: 503–513.

    Article  CAS  Google Scholar 

  7. Kocanova S, Buytaert E, Matroule JY, Piette J, Golab J, de Witte P et al. Induction of heme-oxygenase 1 requires the p38MAPK and PI3K pathways and suppresses apoptotic cell death following hypericin-mediated photodynamic therapy. Apoptosis 2007; 12: 731–741.

    Article  CAS  Google Scholar 

  8. Berberat PO, Dambrauskas Z, Gulbinas A, Giese T, Giese N, Künzli B et al. Inhibition of heme oxygenase-1 increases responsiveness of pancreatic cancer cells to anticancer treatment. Clin Cancer Res 2005; 11: 3790–3798.

    Article  CAS  Google Scholar 

  9. Mayerhofer M, Gleixner KV, Mayerhofer J, Hoermann G, Jaeger E, Aichberger KJ et al. Targeting of heat shock protein 32 (Hsp32)/heme oxygenase-1 (HO-1) in leukemic cells in chronic myeloid leukemia: a novel approach to overcome resistance against imatinib. Blood 2008; 111: 2200–2210.

    Article  CAS  Google Scholar 

  10. Yoshida T, Ishikawa K, Sato M . Degradation of heme by a soluble peptide of heme oxygenase obtained from rat liver microsomes by mild trypsinization. Eur J Biochem 1991; 199: 729–733.

    Article  CAS  Google Scholar 

  11. Lin Q, Weis S, Yang G, Weng YH, Helston R, Rish K et al. Heme oxygenase-1 protein localizes to the nucleus and activates transcription factors important in oxidative stress. J Biol Chem 2007; 282: 20621–20633.

    Article  CAS  Google Scholar 

  12. Sacca P, Meiss R, Casas G, Mazza O, Calvo JC, Navone N et al. Nuclear translocation of haeme oxygenase-1 is associated to prostate cancer. Br J Cancer 2007; 97: 1683–1689.

    Article  CAS  Google Scholar 

  13. Li MY, Yip J, Hsin MK, Mok TS, Wu Y, Underwood MJ et al. Haem oxygenase-1 plays a central role in NNK-mediated lung carcinogenesis. Eur Respir J 2008; 32: 911–923.

    Article  CAS  Google Scholar 

  14. Degese MS, Mendizabal JE, Gandini NA, Gutkind JS, Molinolo A, Hewitt SM et al. Expression of heme oxygenase-1 in non-small cell lung cancer (NSCLC) and its correlation with clinical data. Lung Cancer 2012; 77: 168–175.

    Article  Google Scholar 

  15. Gandini NA, Fermento ME, Salomon DG, Blasco J, Patel V, Gutkind JS et al. Nuclear localization of heme oxygenase-1 is associated with tumor progression of head and neck squamous cell carcinomas. Exp Mol Pathol 2012; 93: 237–245.

    Article  CAS  Google Scholar 

  16. Hwang HW, Lee JR, Chou KY, Suen CS, Hwang MJ, Chen C et al. Oligomerization is crucial for the stability and function of heme oxygenase-1 in the endoplasmic reticulum. J Biol Chem 2009; 284: 22672–22679.

    Article  CAS  Google Scholar 

  17. Fluhrer R, Steiner H, Haass C . Intramembrane proteolysis by signal peptide peptidases: a comparative discussion of GXGD-type aspartyl proteases. J Biol Chem 2009; 284: 13975–13979.

    Article  CAS  Google Scholar 

  18. Converso DP, Taille C, Carreras MC, Jaitovich A, Poderoso JJ, Boczkowski J . HO-1 is located in liver mitochondria and modulates mitochondrial heme content and metabolism. FASEB J 2006; 20: 1236–1238.

    Article  CAS  Google Scholar 

  19. Bindu S, Pal C, Dey S, Goyal M, Alam A, Iqbal MS et al. Translocation of heme oxygenase-1 to mitochondria is a novel cytoprotective mechanism against non-steroidal anti-inflammatory drug-induced mitochondrial oxidative stress, apoptosis, and gastric mucosal injury. J Biol Chem 2011; 286: 39387–39402.

    Article  CAS  Google Scholar 

  20. Wang XM, Kim HP, Nakahira K, Ryter SW, Choi AM . The heme oxygenase-1/carbon monoxide pathway suppresses TLR4 signaling by regulating the interaction of TLR4 with caveolin-1. J Immunol 2009; 182: 3809–3818.

    Article  CAS  Google Scholar 

  21. Suttner DM, Sridhar K, Lee CS, Tomura T, Hansen TN, Dennery PA . Protective effects of transient HO-1 overexpression on susceptibility to oxygen toxicity in lung cells. Am J Physiol 1999; 276: L443–L451.

    CAS  Google Scholar 

  22. Giordano A, Nisoli E, Tonello C, Cancello R, Carruba MO, Cinti S . Expression and distribution of heme oxygenase-1 and -2 in rat brown adipose tissue: the modulatory role of the noradrenergic system. FEBS Lett 2000; 487: 171–175.

    Article  CAS  Google Scholar 

  23. Li Volti G, Ientile R, Abraham NG, Vanella A, Cannavò G, Mazza F et al. Immunocytochemical localization and expression of heme oxygenase-1 in primary astroglial cell cultures during differentiation: effect of glutamate. Biochem Biophys Res Commun 2004; 315: 517–524.

    Article  CAS  Google Scholar 

  24. Lemberg MK, Martoglio B . Requirements for signal peptide peptidase-catalyzed intramembrane proteolysis. Mol Cell 2002; 10: 735–744.

    Article  CAS  Google Scholar 

  25. Chang HT, Kao YL, Wu CM, Fan TC, Lai YK, Huang KL et al. Signal peptide of eosinophil cationic protein upregulates transforming growth factor-alpha expression in human cells. J Cell Biochem 2007; 100: 1266–1275.

    Article  CAS  Google Scholar 

  26. El Hage F, Stroobant V, Vergnon I, Baurain JF, Echchakir H, Lazar V et al. Preprocalcitonin signal peptide generates a cytotoxic T lymphocyte-defined tumor epitope processed by a proteasome-independent pathway. Proc Natl Acad Sci USA 2008; 105: 10119–10124.

    Article  CAS  Google Scholar 

  27. Okamoto K, Moriishi K, Miyamura T, Matsuura Y . Intramembrane proteolysis and endoplasmic reticulum retention of hepatitis C virus core protein. J Virol 2004; 78: 6370–6380.

    Article  CAS  Google Scholar 

  28. Targett-Adams P, Schaller T, Hope G, Lanford RE, Lemon SM, Martin A et al. Signal peptide peptidase cleavage of GB virus B core protein is required for productive infection in vivo. J Biol Chem 2006; 281: 29221–29227.

    Article  CAS  Google Scholar 

  29. Erez E, Fass D, Bibi E . How intramembrane proteases bury hydrolytic reactions in the membrane. Nature 2009; 459: 371–378.

    Article  CAS  Google Scholar 

  30. Pace CN, Scholtz JM . A helix propensity scale based on experimental studies of peptides and proteins. Biophys J 1998; 75: 422–427.

    Article  CAS  Google Scholar 

  31. Okamoto K, Mori Y, Komoda Y, Okamoto T, Okochi M, Takeda M et al. Intramembrane processing by signal peptide peptidase regulates the membrane localization of hepatitis C virus core protein and viral propagation. J Virol 2008; 82: 8349–8361.

    Article  CAS  Google Scholar 

  32. Collinson EJ, Wimmer-Kleikamp S, Gerega SK, Yang YH, Parish CR, Dawes IW et al. The yeast homolog of heme oxygenase-1 affords cellular antioxidant protection via the transcriptional regulation of known antioxidant genes. J Biol Chem 2011; 286: 2205–2214.

    Article  CAS  Google Scholar 

  33. Lin PH, Lan WM, Chau LY . TRC8 suppresses tumorigenesis through targeting heme oxygenase-1 for ubiquitination and degradation. Oncogene 2013; 32: 2325–2334.

    Article  CAS  Google Scholar 

  34. Wen W, Meinkoth JL, Tsien RY, Taylor SS . Identification of a signal for rapid export of proteins from the nucleus. Cell 1995; 82: 463–473.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by a grant from the National Science Council of Taiwan (NSC-100-2320-B-001-010-MY3). We thank the National Center for Genome Medicine at Academia Sinica, Taiwan for polymerase chain reaction of short-tandem repeat sequences analysis, and the IBMS proteomics core for LC-MS analysis.

Author information

Authors and Affiliations

  1. Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan

    F-F Hsu, Y-J Sun, M-T Chiang, W-M Lan, F-A Li & L-Y Chau

  2. Graduate Institute of Clinical Medicine, Taipei Medical University, Taipei, Taiwan

    C-T Yeh

  3. Cancer Center, Shuang Ho Hospital, Taipei Medical University, Taipei, Taiwan

    C-T Yeh & W-H Lee

Authors
  1. F-F Hsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C-T Yeh
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Y-J Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M-T Chiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. W-M Lan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. F-A Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. W-H Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. L-Y Chau
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to L-Y Chau.

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

Hsu, FF., Yeh, CT., Sun, YJ. et al. Signal peptide peptidase-mediated nuclear localization of heme oxygenase-1 promotes cancer cell proliferation and invasion independent of its enzymatic activity. Oncogene 34, 2360–2370 (2015). https://doi.org/10.1038/onc.2014.166

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2014.166

This article is cited by

Search

Advanced search

Quick links