Molecular and Cellular Biochemistry

, Volume 301, Issue 1–2, pp 115–122 | Cite as

Association of LKB1 with a WD-repeat protein WDR6 is implicated in cell growth arrest and p27Kip1 induction

Article

Abstract

Germline mutations of the serine/threonine kinase LKB1 (also known as STK11) lead to Peutz–Jeghers syndrome (PJS) that is associated with increased incidence of malignant cancers. However, the tumor suppressor function of LKB1 has not been fully elucidated. We applied yeast two-hybrid screening and identified that a novel WD-repeat protein WDR6 was able to interact with LKB1. Immunofluorescence staining revealed that WDR6 was localized in cytoplasm, similar to the localization of LKB1. Expression of LKB1 was able to inhibit colony formation of Hela cells. Interestingly, coexpression of WDR6 with LKB1 enhanced the inhibitory effect of LKB1 on Hela cell proliferation. Consistently, WDR6 was able to synergize with LKB1 in cell cycle G1 arrest in Hela cells. Coexpression of WDR6 and LKB1 was able to induce a cyclin-dependent kinase (CDK) inhibitor p27Kip1. Furthermore, the stimulatory effect of LKB1 on p27Kip1 promoter activity was significantly elevated by coexpression with WDR6. Collectively, these results provided initial evidence that WDR6 is implicated in the cell growth inhibitory pathway of LKB1 via regulation of p27Kip1.

Keywords

LKB1 WDR6 Cell cycle Tumor suppressor p27Kip1 Peutz–Jeghers syndrome WD-repeat 

Abbreviations

AMPK

AMP-activated protein kinase

CDK

Cyclin-dependent kinase

CKI

Cyclin-dependent kinase inhibitors

EGFP

Enhanced green fluorescence protein

G proteins β subunit

Notes

Acknowledgments

We wish to thank Dr. Mackintosh at Dundee, UK for providing the human LKB1 plasmid. This work was supported by research grants from Chinese Academy of Sciences (One Hundred Talents program), National Natural Science Foundation of China (30470870 and 30588002), and Science & Technology Commission of Shanghai Municipality (05DJ14009 and 04DZ14007) to YC.

References

  1. 1.
    Alessi DR, Sakamoto K, Bayascas JR (2006) Lkb1-dependent signaling pathways. Annu Rev Biochem 75:137–163PubMedCrossRefGoogle Scholar
  2. 2.
    Marignani PA (2005) LKB1, the multitasking tumour suppressor kinase. J Clin Pathol 58:15–19PubMedCrossRefGoogle Scholar
  3. 3.
    Hemminki A, Markie D, Tomlinson I, Avizienyte E, Roth S, Loukola A, Bignell G, Warren W, Aminoff M, Hoglund P, Jarvinen H, Kristo P, Pelin K, Ridanpaa M, Salovaara R, Toro T, Bodmer W, Olschwang S, Olsen AS, Stratton MR, de la Chapelle A, Aaltonen LA (1998) A serine/threonine kinase gene defective in Peutz–Jeghers syndrome. Nature 391:184–187PubMedCrossRefGoogle Scholar
  4. 4.
    Jenne DE, Reimann H, Nezu J, Friedel W, Loff S, Jeschke R, Muller O, Back W, Zimmer M (1998) Peutz–Jeghers syndrome is caused by mutations in a novel serine threonine kinase. Nat Genet 18:38–43PubMedCrossRefGoogle Scholar
  5. 5.
    Giardiello FM, Welsh SB, Hamilton SR, Offerhaus GJ, Gittelsohn AM, Booker SV, Krush AJ, Yardley JH, Luk GD (1987) Increased risk of cancer in the Peutz–Jeghers syndrome. N Engl J Med 316:1511–1514PubMedCrossRefGoogle Scholar
  6. 6.
    Tiainen M, Ylikorkala A, Makela TP (1999) Growth suppression by Lkb1 is mediated by a G(1) cell cycle arrest. Proc Natl Acad Sci USA 96:9248–9251PubMedCrossRefGoogle Scholar
  7. 7.
    Tiainen M, Vaahtomeri K, Ylikorkala A, Makela TP (2002) Growth arrest by the LKB1 tumor suppressor: induction of p21(WAF1/CIP1). Hum Mol Genet 11:1497–1504PubMedCrossRefGoogle Scholar
  8. 8.
    Marignani PA, Kanai F, Carpenter CL (2001) LKB1 associates with Brg1 and is necessary for Brg1-induced growth arrest. J Biol Chem 276:32415–32418PubMedCrossRefGoogle Scholar
  9. 9.
    Ylikorkala A, Rossi DJ, Korsisaari N, Luukko K, Alitalo K, Henkemeyer M, Makela TP (2001) Vascular abnormalities and deregulation of VEGF in Lkb1-deficient mice. Science 293:1323–1326PubMedCrossRefGoogle Scholar
  10. 10.
    Miyoshi H, Nakau M, Ishikawa TO, Seldin MF, Oshima M, Taketo MM (2002) Gastrointestinal hamartomatous polyposis in Lkb1 heterozygous knockout mice. Cancer Res 62:2261–2266PubMedGoogle Scholar
  11. 11.
    Carling D (2006) LKB1:a sweet side to Peutz–Jeghers syndrome? Trends Mol Med 12:144–147PubMedCrossRefGoogle Scholar
  12. 12.
    Hawley SA, Boudeau J, Reid JL, Mustard KJ, Udd L, Makela TP, Alessi DR, Hardie DG (2003) Complexes between the LKB1 tumor suppressor, STRAD alpha/beta and MO25 alpha/beta are upstream kinases in the AMP-activated protein kinase cascade. J Biol 2:28PubMedCrossRefGoogle Scholar
  13. 13.
    Woods A, Johnstone SR, Dickerson K, Leiper FC, Fryer LG, Neumann D, Schlattner U, Wallimann T, Carlson M, Carling D (2003) LKB1 is the upstream kinase in the AMP-activated protein kinase cascade. Curr Biol 13:2004–2008PubMedCrossRefGoogle Scholar
  14. 14.
    Lizcano JM, Goransson O, Toth R, Deak M, Morrice NA, Boudeau J, Hawley SA, Udd L, Makela TP, Hardie DG, Alessi DR (2004) LKB1 is a master kinase that activates 13 kinases of the AMPK subfamily, including MARK/PAR-1. Embo J 23:833–843PubMedCrossRefGoogle Scholar
  15. 15.
    Shaw RJ, Kosmatka M, Bardeesy N, Hurley RL, Witters LA, DePinho RA, Cantley LC (2004) The tumor suppressor LKB1 kinase directly activates AMP-activated kinase and regulates apoptosis in response to energy stress. Proc Natl Acad Sci USA 101:3329–3335PubMedCrossRefGoogle Scholar
  16. 16.
    Musi N (2006) AMP-activated protein kinase and type 2 diabetes. Curr Med Chem 13:583–589PubMedCrossRefGoogle Scholar
  17. 17.
    Sakamoto K, McCarthy A, Smith D, Green KA, Grahame Hardie D, Ashworth A, Alessi DR (2005) Deficiency of LKB1 in skeletal muscle prevents AMPK activation and glucose uptake during contraction. Embo J 24:1810–1820PubMedCrossRefGoogle Scholar
  18. 18.
    Shaw RJ, Lamia KA, Vasquez D, Koo SH, Bardeesy N, Depinho RA, Montminy M, Cantley LC (2005) The kinase LKB1 mediates glucose homeostasis in liver and therapeutic effects of metformin. Science 310:1642–1646PubMedCrossRefGoogle Scholar
  19. 19.
    Godbey WT, Wu KK, Hirasaki GJ, Mikos AG (1999) Improved packing of poly(ethylenimine)/DNA complexes increases transfection efficiency. Gene Ther 6:1380–1388PubMedCrossRefGoogle Scholar
  20. 20.
    Sun X, Han R, Wang Z, Chen Y (2006) Regulation of adiponectin receptors in hepatocytes by the peroxisome proliferator-activated receptor-gamma agonist rosiglitazone. Diabetologia 49:1303–1310PubMedCrossRefGoogle Scholar
  21. 21.
    Li D, Burch P, Gonzalez O, Kashork CD, Shaffer LG, Bachinski LL, Roberts R (2000) Molecular cloning, expression analysis, and chromosome mapping of WDR6, a novel human WD-repeat gene. Biochem Biophys Res Commun 274:117–123PubMedCrossRefGoogle Scholar
  22. 22.
    Slingerland J, Pagano M (2000) Regulation of the cdk inhibitor p27 and its deregulation in cancer. J Cell Physiol 183:10–17PubMedCrossRefGoogle Scholar
  23. 23.
    Sgambato A, Cittadini A, Faraglia B, Weinstein IB (2000) Multiple functions of p27(Kip1) and its alterations in tumor cells:a review. J Cell Physiol 183:18–27PubMedCrossRefGoogle Scholar
  24. 24.
    Shen Z, Wen XF, Lan F, Shen ZZ, Shao ZM (2002) The tumor suppressor gene LKB1 is associated with prognosis in human breast carcinoma. Clin Cancer Res 8:2085–2090PubMedGoogle Scholar
  25. 25.
    Setogawa T, Shinozaki-Yabana S, Masuda T, Matsuura K, Akiyama T (2006) The tumor suppressor LKB1 induces p21 expression in collaboration with LMO4, GATA-6, and Ldb1. Biochem Biophys Res Commun 343:1186–1190PubMedCrossRefGoogle Scholar
  26. 26.
    Neer EJ, Schmidt CJ, Nambudripad R, Smith TF (1994) The ancient regulatory-protein family of WD-repeat proteins. Nature 371:297–300PubMedCrossRefGoogle Scholar
  27. 27.
    Smith TF, Gaitatzes C, Saxena K, Neer EJ (1999) The WD repeat: a common architecture for diverse functions. Trends Biochem Sci 24:181–185PubMedCrossRefGoogle Scholar
  28. 28.
    Li D, Roberts R (2001) WD-repeat proteins: structure characteristics, biological function, and their involvement in human diseases. Cell Mol Life Sci 58:2085–2097PubMedCrossRefGoogle Scholar
  29. 29.
    Wall MA, Coleman DE, Lee E, Iniguez-Lluhi JA, Posner BA, Gilman AG, Sprang SR (1995) The structure of the G protein heterotrimer Gi alpha 1 beta 1 gamma 2. Cell 83:1047–1058PubMedCrossRefGoogle Scholar
  30. 30.
    Sondek J, Bohm A, Lambright DG, Hamm HE, Sigler PB (1996) Crystal structure of a G-protein beta gamma dimer at 2.1A resolution. Nature 379:369–374PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  1. 1.Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences (SIBS)Chinese Academy of Sciences, Graduate School of the Chinese Academy of SciencesShanghaiChina

Personalised recommendations