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:

Nuclear import of LASP-1 is regulated by phosphorylation and dynamic protein–protein interactions

Oncogene volume 32, pages 2107–2113 (2013)Cite this article

Subjects

Abstract

LASP-1 is a multidomain protein predominantly localized at focal contacts, where it regulates cytoskeleton dynamics and cell migration. However, in different tumor entities, a nuclear LASP-1 accumulation is observed, thought to have an important role in cancer progression. Until now, the molecular mechanisms that control LASP-1 nuclear import were not elucidated. Here, we identified a novel LASP-1-binding partner, zona occludens protein 2 (ZO-2), and established its role in the signal transduction pathway of LASP-1 nucleo-cytoplasmatic shuttling. Phosphorylation of LASP-1 by PKA at serine 146 induces translocation of the LASP-1/ZO-2 complex from the cytoplasm to the nucleus. Interaction occurs within the carboxyterminal proline-rich motif of ZO-2 and the SH3 domain in LASP-1. In situ proximity ligation assay confirmed the direct binding between LASP-1 and ZO-2 and visualized the shuttling. Nuclear export is mediated by Crm-1 and a newly identified nuclear export signal in LASP-1. Finally, dephosphorylation of LASP-1 by phosphatase PP2B is suggested to relocalize the protein back to focal contacts. In summary, we define a new pathway for LASP-1 in tumor progression.

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

Accession codes

Accessions

GenBank/EMBL/DDBJ

References

  1. Tomasetto C, Regnier C, Moog-Lutz C, Mattei MG, Chenard MP, Lidereau R et al. Identification of four novel human genes amplified and overexpressed in breast carcinoma and localized to the q11-q21.3 region of chromosome 17. Genomics 1995; 28: 367–376.

    Article  CAS  Google Scholar 

  2. Grunewald TG, Butt E . The LIM and SH3 domain protein family: structural proteins or signal transducers or both? Mol Cancer 2008; 7: 31.

    Article  Google Scholar 

  3. Grunewald TG, Kammerer U, Schulze E, Schindler D, Honig A, Zimmer M et al. Silencing of LASP-1 influences zyxin localization, inhibits proliferation and reduces migration in breast cancer cells. Exp Cell Res 2006; 312: 974–982.

    Article  CAS  Google Scholar 

  4. Grunewald TG, Kammerer U, Winkler C, Schindler D, Sickmann A, Honig A et al. Overexpression of LASP-1 mediates migration and proliferation of human ovarian cancer cells and influences zyxin localisation. Br J Cancer 2007; 96: 296–305.

    Article  CAS  Google Scholar 

  5. Dimova I, Orsetti B, Negre V, Rouge C, Ursule L, Lasorsa L et al. Genomic markers for ovarian cancer at chromosomes 1, 8 and 17 revealed by array CGH analysis. Tumori 2009; 95: 357–366.

    Article  CAS  Google Scholar 

  6. Zhao L, Wang H, Liu C, Liu Y, Wang X, Wang S et al. Promotion of colorectal cancer growth and metastasis by the LIM and SH3 domain protein 1. Gut 2010; 59: 1226–1235.

    Article  CAS  Google Scholar 

  7. Traenka C, Remke M, Korshunov A, Bender S, Hielscher T, Northcott PA et al. Role of LIM and SH3 protein 1 (LASP1) in the metastatic dissemination of medulloblastoma. Cancer Res 2010; 70: 8003–8014.

    Article  CAS  Google Scholar 

  8. Nakagawa H, Terasaki AG, Suzuki H, Ohashi K, Miyamoto S . Short-term retention of actin filament binding proteins on lamellipodial actin bundles. FEBS lett 2006; 580: 3223–3228.

    Article  CAS  Google Scholar 

  9. Grunewald TG, Kammerer U, Kapp M, Eck M, Dietl J, Butt E et al. Nuclear localization and cytosolic overexpression of LASP-1 correlates with tumor size and nodal-positivity of human breast carcinoma. BMC cancer 2007; 7: 198.

    Article  Google Scholar 

  10. Zhang H, Chen X, Bollag WB, Bollag RJ, Sheehan DJ, Chew CS . Lasp1 gene disruption is linked to enhanced cell migration and tumor formation. Physiol Genomics 2009; 38: 372–385.

    Article  CAS  Google Scholar 

  11. Chew CS, Chen X, Parente JA, Tarrer S, Okamoto C, Qin HY . Lasp-1 binds to non-muscle F-actin in vitro and is localized within multiple sites of dynamic actin assembly in vivo. J Cell Sci 2002; 115: 4787–4799.

    Article  CAS  Google Scholar 

  12. Butt E, Gambaryan S, Gottfert N, Galler A, Marcus K, Meyer HE . Actin binding of human LIM and SH3 protein is regulated by cGMP- and cAMP-dependent protein kinase phosphorylation on serine 146. J Biol Chem 2003; 278: 15601–15607.

    Article  CAS  Google Scholar 

  13. Lin YH, Park ZY, Lin D, Brahmbhatt AA, Rio MC, Yates JR et al. Regulation of cell migration and survival by focal adhesion targeting of Lasp-1. J Cell Biol 2004; 165: 421–432.

    Article  CAS  Google Scholar 

  14. Traenka J, Hauck CR, Lewandrowski U, Sickmann A, Gambaryan S, Thalheimer P et al. Integrin-dependent translocation of LASP-1 to the cytoskeleton of activated platelets correlates with LASP-1 phosphorylation at tyrosine 171 by Src-kinase. Thromb Haemost 2009; 102: 520–528.

    Article  CAS  Google Scholar 

  15. Frietsch JJ, Grunewald TG, Jasper S, Kammerer U, Herterich S, Kapp M et al. Nuclear localisation of LASP-1 correlates with poor long-term survival in female breast cancer. Br J Cancer 2010; 102: 1645–1653.

    Article  CAS  Google Scholar 

  16. Kudo N, Wolff B, Sekimoto T, Schreiner EP, Yoneda Y, Yanagida M et al. Leptomycin B inhibition of signal-mediated nuclear export by direct binding to CRM1. Exper Cell Res 1998; 242: 540–547.

    Article  CAS  Google Scholar 

  17. Lerea KM . Thrombin-induced effects are selectively inhibited following treatment of intact human platelets with okadaic acid. Biochemistry 1991; 30: 6819–6824.

    Article  CAS  Google Scholar 

  18. Kutay U, Guttinger S . Leucine-rich nuclear-export signals: born to be weak. Trends Cell Biol 2005; 15: 121–124.

    Article  CAS  Google Scholar 

  19. Li B, Zhuang L, Trueb B . Zyxin interacts with the SH3 domains of the cytoskeletal proteins LIM-nebulette and Lasp-1. The J Biol Chem 2004; 279: 20401–20410.

    Article  CAS  Google Scholar 

  20. Chew CS, Chen X, Bollag RJ, Isales CM, Ding KH, Zhang H . Targeted disruption of the Lasp1 gene is linked to increases in histamine-stimulated gastric HCl secretion. Am J Physiol Gastrointest Liver Physiol 2008; 288: G376–G381.

    Article  Google Scholar 

  21. Betanzos A, Huerta M, Lopez-Bayghen E, Azuara E, Amerena J, Gonzalez-Mariscal L . The tight junction protein ZO-2 associates with Jun, Fos and C/EBP transcription factors in epithelial cells. Exper Cell Res 2004; 292: 51–66.

    Article  CAS  Google Scholar 

  22. Keicher C, Gambaryan S, Schulze E, Marcus K, Meyer HE, Butt E . Phosphorylation of mouse LASP-1 on threonine 156 by cAMP- and cGMP-dependent protein kinase. Biochem Biophys Res Commun 2004; 324: 308–316.

    Article  CAS  Google Scholar 

  23. Ferraro E, Peluso D, Via A, Ausiello G, Helmer-Citterich M . SH3-hunter: discovery of SH3 domain interaction sites in proteins. Nucleic Acids Res 2007; 35: W451–W454.

    Article  Google Scholar 

  24. Soderberg O, Gullberg M, Jarvius M, Ridderstrale K, Leuchowius KJ, Jarvius J et al. Direct observation of individual endogenous protein complexes in situ by proximity ligation. Nat Methods 2006; 3: 995–1000.

    Article  Google Scholar 

  25. Chiyomaru T, Enokida H, Kawakami K, Tatarano S, Uchida Y, Kawahara K et al. Functional role of LASP1 in cell viability and its regulation by microRNAs in bladder cancer. Urol Oncol 2010, (e-pub ahead of print).

  26. Viticchie G, Lena AM, Latina A, Formosa A, Gregersen LH, Lund AH et al. MiR-203 controls proliferation, migration and invasive potential of prostate cancer cell lines. Cell Cycle 10: 1121–1131.

    Article  Google Scholar 

  27. Call GS, Chung JY, Davis JA, Price BD, Primavera TS, Thomson NC et al. Zyxin phosphorylation at serine 142 modulates the zyxin head-tail interaction to alter cell-cell adhesion. Biochem Biophys Res Commun 404: 780–784.

    Article  CAS  Google Scholar 

  28. Merkle D, Hoffmann R . Roles of cAMP and cAMP-dependent protein kinase in the progression of prostate cancer: cross-talk with the androgen receptor. Cell Signal 2011; 23: 507–515.

    Article  CAS  Google Scholar 

  29. Traweger A, Fuchs R, Krizbai IA, Weiger TM, Bauer HC, Bauer H . The tight junction protein ZO-2 localizes to the nucleus and interacts with the heterogeneous nuclear ribonucleoprotein scaffold attachment factor-B. J Biol Chem 2003; 278: 2692–2700.

    Article  CAS  Google Scholar 

  30. Balda MS, Matter K . Tight junctions and the regulation of gene expression. Biochim Biophys Acta 2009; 1788: 761–767.

    Article  CAS  Google Scholar 

  31. Huerta M, Munoz R, Tapia R, Soto-Reyes E, Ramirez L, Recillas-Targa F et al. Cyclin D1 is transcriptionally down-regulated by ZO-2 via an E box and the transcription factor c-Myc. Mol Biol Cell 2007; 18: 4826–4836.

    Article  CAS  Google Scholar 

  32. Oka T, Remue E, Meerschaert K, Vanloo B, Boucherie C, Gfeller D et al. Functional complexes between YAP2 and ZO-2 are PDZ domain-dependent, and regulate YAP2 nuclear localization and signalling. Biochem J 432: 461–472.

    Article  CAS  Google Scholar 

  33. Hammarstrom A, Berndt KD, Sillard R, Adermann K, Otting G . Solution structure of a naturally-occurring zinc-peptide complex demonstrates that the N-terminal zinc-binding module of the Lasp-1 LIM domain is an independent folding unit. Biochemistry 1996; 35: 12723–12732.

    Article  CAS  Google Scholar 

  34. Zheng B, Han M, Bernier M, Wen JK . Nuclear actin and actin-binding proteins in the regulation of transcription and gene expression. FEBS J 2009; 276: 2669–2685.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Dr Viacheslav Nikolaev for his help with confocal microscopy. This work was supported by grants from the DFG (BU 740/6-2 to EB, RE 3183/1-1 to CR, SFB 688 A2 to SG) the Innovative Medizinische Forschung, University Hospital Münster (KR110904 to JK) and the Ministerium für Innovation, Wissenschaft und Forschung des Landes Nordrhein-Westfalen (UL and AS).

Author information

Authors and Affiliations

  1. Institute of Clinical Biochemistry and Pathobiochemistry, University Clinic of Wuerzburg, Wuerzburg, Germany

    S Mihlan, C Reiß, P Thalheimer, S Herterich, S Gaetzner & E Butt

  2. Department of Internal Medicine D, Division for Molecular Nephrology, University Hospital Muenster, Muenster, Germany

    J Kremerskothen & H J Pavenstädt

  3. Department of Bioanalytics, Leibniz-Institut fuer Analytische Wissenschaften–ISAS–e.V., Dortmund, Germany

    U Lewandrowski & A Sickmann

  4. Medical Proteome Center, Ruhr-University, Bochum, Germany

    A Sickmann

Authors
  1. S Mihlan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C Reiß
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. P Thalheimer
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S Herterich
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. S Gaetzner
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. J Kremerskothen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. H J Pavenstädt
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. U Lewandrowski
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. A Sickmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. E Butt
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to E Butt.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies the paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mihlan, S., Reiß, C., Thalheimer, P. et al. Nuclear import of LASP-1 is regulated by phosphorylation and dynamic protein–protein interactions. Oncogene 32, 2107–2113 (2013). https://doi.org/10.1038/onc.2012.216

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2012.216

Keywords

This article is cited by

Search

Advanced search

Quick links