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Cytoplasmic PML function in TGF-β signalling

Nature volume 431, pages 205–211 (2004)Cite this article

Abstract

Transforming growth factor β (TGF-β) is a pluripotent cytokine that controls key tumour suppressive functions1,2,3, but cancer cells are often unresponsive to it1,4. The promyelocytic leukaemia (PML) tumour suppressor of acute promyelocytic leukaemia (APL) accumulates in the PML nuclear body, but cytoplasmic PML isoforms of unknown function have also been described5,6. Here we show that cytoplasmic Pml is an essential modulator of TGF-β signalling. Pml-null primary cells are resistant to TGF-β-dependent growth arrest, induction of cellular senescence and apoptosis. These cells also have impaired phosphorylation and nuclear translocation of the TGF-β signalling proteins Smad2 and Smad3, as well as impaired induction of TGF-β target genes. Expression of cytoplasmic Pml is induced by TGF-β. Furthermore, cytoplasmic PML physically interacts with Smad2/3 and SARA (Smad anchor for receptor activation) and is required for association of Smad2/3 with SARA and for the accumulation of SARA and TGF-β receptor in the early endosome. The PML–RARα oncoprotein of APL can antagonize cytoplasmic PML function and APL cells have defects in TGF-β signalling similar to those observed in Pml-null cells. Our findings identify cytoplasmic PML as a critical TGF-β regulator, and further implicate deregulated TGF-β signalling in cancer pathogenesis.

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Figure 1: Pml is essential for TGF-β1 transcriptional and biological functions.
Figure 4: Aberrant TGF-β1 signalling in APL.
Figure 2: cPML interacts physically and functionally with Smad2/3 and SARA.
Figure 3: PML is required for TβRI/II and SARA localization in the early endosome.

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References

  1. Derynck, R., Akhurst, R. J. & Balmain, A. TGF-β signaling in tumor suppression and cancer progression. Nature Genet. 29, 117–129 (2001)

    Article  CAS  PubMed  Google Scholar 

  2. Siegel, P. M. & Massague, J. Cytostatic and apoptotic actions of TGF-β in homeostasis and cancer. Nature Rev. Cancer 3, 807–820 (2003)

    Article  CAS  Google Scholar 

  3. Attisano, L. & Wrana, J. L. Signal transduction by the TGF-β superfamily. Science 296, 1646–1647 (2002)

    Article  ADS  CAS  PubMed  Google Scholar 

  4. Kim, S. J. & Letterio, J. Transforming growth factor-β signaling in normal and malignant hematopoiesis. Leukemia 17, 1731–1737 (2003)

    Article  CAS  PubMed  Google Scholar 

  5. Jensen, K., Shiels, C. & Freemont, P. S. PML protein isoforms and the RBCC/TRIM motif. Oncogene 20, 7223–7233 (2001)

    Article  CAS  PubMed  Google Scholar 

  6. Fagioli, M. et al. Alternative splicing of PML transcripts predicts coexpression of several carboxy-terminally different protein isoforms. Oncogene 7, 1083–1091 (1992)

    CAS  PubMed  Google Scholar 

  7. Melnick, A. & Licht, J. D. Deconstructing a disease: RARα, its fusion partners, and their roles in the pathogenesis of acute promyelocytic leukemia. Blood 93, 3167–3215 (1999)

    CAS  PubMed  Google Scholar 

  8. Salomoni, P. & Pandolfi, P. P. The role of PML in tumor suppression. Cell 108, 165–170 (2002)

    Article  CAS  PubMed  Google Scholar 

  9. Kastner, P. et al. Structure, localization and transcriptional properties of two classes of retinoic acid receptor α fusion proteins in acute promyelocytic leukemia (APL): structural similarities with a new family of oncoproteins. EMBO J. 11, 629–642 (1992)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Ferbeyre, G. et al. PML is induced by oncogenic ras and promotes premature senescence. Genes Dev. 14, 2015–2027 (2000)

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Pearson, M. et al. PML regulates p53 acetylation and premature senescence induced by oncogenic Ras. Nature 406, 207–210 (2000)

    Article  ADS  CAS  PubMed  Google Scholar 

  12. Katakura, Y., Nakata, E., Miura, T. & Shirahata, S. Transforming growth factor βtriggers two independent-senescence programs in cancer cells. Biochem. Biophys. Res. Commun. 255, 110–115 (1999)

    Article  CAS  PubMed  Google Scholar 

  13. Tsukazaki, T., Chiang, T. A., Davison, A. F., Attisano, L. & Wrana, J. L. SARA, a FYVE domain protein that recruits Smad2 to the TGFβ receptor. Cell 95, 779–791 (1998)

    Article  CAS  PubMed  Google Scholar 

  14. Di Guglielmo, G. M., Le Roy, C., Goodfellow, A. F. & Wrana, J. L. Distinct endocytic pathways regulate TGF-β receptor signalling and turnover. Nature Cell Biol. 5, 410–421 (2003)

    Article  CAS  PubMed  Google Scholar 

  15. Panopoulou, E. et al. Early endosomal regulation of Smad-dependent signaling in endothelial cells. J. Biol. Chem. 277, 18046–18052 (2002)

    Article  CAS  PubMed  Google Scholar 

  16. Hayes, S., Chawla, A. & Corvera, S. TGF βreceptor internalization into EEA1-enriched early endosomes: role in signaling to Smad2. J. Cell. Biol. 158, 1239–1249 (2002)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Testa, U. et al. PML/RAR α + U937 mutant and NB4 cell lines: retinoic acid restores the monocytic differentiation response to vitamin D3. Cancer Res. 54, 4508–4515 (1994)

    CAS  PubMed  Google Scholar 

  18. Grignani, F. et al. The acute promyelocytic leukemia-specific PML–RAR α fusion protein inhibits differentiation and promotes survival of myeloid precursor cells. Cell 74, 423–431 (1993)

    Article  CAS  PubMed  Google Scholar 

  19. Testa, U. et al. Transforming growth factor-β potentiates vitamin D3-induced terminal monocytic differentiation of human leukemic cell lines. J. Immunol. 150, 2418–2430 (1993)

    CAS  PubMed  Google Scholar 

  20. Zhu, J., Chen, Z., Lallemand-Breitenbach, V. & de The, H. How acute promyelocytic leukaemia revived arsenic. Nature Rev. Cancer 2, 705–713 (2002)

    Article  CAS  Google Scholar 

  21. Turelli, P. et al. Cytoplasmic recruitment of INI1 and PML on incoming HIV preintegration complexes: interference with early steps of viral replication. Mol. Cell 7, 1245–1254 (2001)

    Article  CAS  PubMed  Google Scholar 

  22. Cordenonsi, M. et al. Links between tumor suppressors: p53 is required for TGF-β gene responses by cooperating with Smads. Cell 113, 301–314 (2003)

    Article  CAS  PubMed  Google Scholar 

  23. Gurrieri, C. et al. Loss of the tumor suppressor PML in human cancers of multiple histologic origins. J. Natl Cancer Inst. 96, 269–279 (2004)

    Article  CAS  PubMed  Google Scholar 

  24. Howe, J. R. et al. Mutations in the SMAD4/DPC4 gene in juvenile polyposis. Science 280, 1086–1088 (1998)

    Article  ADS  CAS  PubMed  Google Scholar 

  25. Miyaki, M. et al. Higher frequency of Smad4 gene mutation in human colorectal cancer with distant metastasis. Oncogene 18, 3098–3103 (1999)

    Article  CAS  PubMed  Google Scholar 

  26. Wang, Z. G. et al. Role of PML in cell growth and the retinoic acid pathway. Science 279, 1547–1551 (1998)

    Article  ADS  CAS  PubMed  Google Scholar 

  27. Kang, H. Y. et al. From transforming growth factor-β signaling to androgen action: identification of Smad3 as an androgen receptor coregulator in prostate cancer cells. Proc. Natl Acad. Sci. USA 98, 3018–3023 (2001)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  28. Lin, H. K., Wang, L., Hu, Y. C., Altuwaijri, S. & Chang, C. Phosphorylation-dependent ubiquitylation and degradation of androgen receptor by Akt require Mdm2 E3 ligase. EMBO J. 21, 4037–4048 (2002)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Kurokawa, M. et al. The oncoprotein Evi-1 represses TGF-β signalling by inhibiting Smad3. Nature 394, 92–96 (1998)

    Article  ADS  CAS  PubMed  Google Scholar 

  30. Escola, J. M., Grivel, J. C., Chavrier, P. & Gorvel, J. P. Different endocytic compartments are involved in the tight association of class II molecules with processed hen egg lysozyme and ribonuclease A in B cells. J. Cell Sci. 108, 2337–2345 (1995)

    CAS  PubMed  Google Scholar 

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Acknowledgements

We are grateful to C. Chang, K. S. Chang, H. Y. Kang, Y. B. Kang, S. W. Lowe, P. G. Pelicci, M. Reiss, T. Sollner and J. L. Wrana for reagents and advice. Special thanks to all the members of the Pandolfi laboratory for comments and discussion. We also thank T. Merghoub and I. Guernah for technical help, R. Hobbs and L. DiSantis for critical reading and preparation of the manuscript. This work was supported by National Institutes of Health grants to P.P.P.

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Authors and Affiliations

  1. Cancer Biology and Genetics Program, Department of Pathology, Memorial Sloan-Kettering Cancer Center, Sloan-Kettering Institute, 1275 York Avenue, New York, New York, 10021, USA

    Hui-Kuan Lin, Stephan Bergmann & Pier Paolo Pandolfi

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  1. Hui-Kuan Lin
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  2. Stephan Bergmann
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  3. Pier Paolo Pandolfi
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Correspondence to Pier Paolo Pandolfi.

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Supplementary information

Supplementary Methods (DOC 20 kb)

Supplementary Figure Legends (DOC 24 kb)

Supplementary Figure 1

cPML is induced by TGF-β1 and exhibits a punctate cytoplasmic localization pattern. (PPT 790 kb)

Supplementary Figure 2

Role of cPml in TGF-β1 signal transduction. (PPT 1480 kb)

Supplementary Figure 3

Determination of the cPML and Smad3 regions required for their physical interaction. (PPT 407 kb)

Supplementary Figure 4

cPML, SARA, and Smad3 colocalize in cytoplasmic region. (PPT 1186 kb)

Supplementary Figure 5

Colocalization of Smad3 and SARA is impaired in Pml-/- MEFs. (PPT 222 kb)

Supplementary Figure 6

Localization of endogenous TbRI to the early endosome is impaired in Pml-/- MEFs. (PPT 763 kb)

Supplementary Figure 7

Endogenous cPml colocalizes with EEA1. (PPT 354 kb)

Supplementary Figure 8

cPML interacts with TβRI and TβRII. (PPT 196 kb)

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Lin, HK., Bergmann, S. & Pandolfi, P. Cytoplasmic PML function in TGF-β signalling. Nature 431, 205–211 (2004). https://doi.org/10.1038/nature02783

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