, Volume 27, Issue 6, pp 1203–1216 | Cite as

Beryllium is an inhibitor of cellular GSK-3β that is 1,000-fold more potent than lithium

  • Swapna R. Mudireddy
  • Ataur Rahman Mohammed Abdul
  • Priyatham Gorjala
  • Ronald K. Gary


Glycogen synthase kinase 3β (GSK-3β) is a key regulator in signaling networks that control cell proliferation, metabolism, development, and other processes. Lithium chloride is a GSK-3 family inhibitor that has been a mainstay of in vitro and in vivo studies for many years. Beryllium salt has the potential to act as a lithium-like inhibitor of GSK-3, but it is not known whether this agent is effective under physiologically relevant conditions. Here we show that BeSO4 inhibits endogenous GSK-3β in cultured human cells. Exposure to 10 µM Be2+ produced a decrease in GSK-3β kinase activity that was comparable to that produced by 10 mM Li+, indicating that beryllium is about 1,000-fold more potent than the classical inhibitor when treating intact cells. There was a statistically significant dose-dependent reduction in specific activity of GSK-3β immunoprecipitated from cells that had been treated with either agent. Lithium inhibited GSK-3β kinase activity directly, and it also caused GSK-3β in cells to become phosphorylated at serine-9 (Ser-9), a post-translational modification that occurs as part of a well-known positive feedback loop that suppresses the kinase activity. Beryllium also inhibited the kinase directly, but unlike lithium it had little effect on Ser-9 phosphorylation in the cell types tested, suggesting that alternative modes of feedback inhibition may be elicited by this agent. These results indicate that beryllium, like lithium, can induce perturbations in the GSK-3β signaling network of treated cells.


Beryllium sulfate Lithium chloride Glycogen synthase kinase Phosphorylation 



We thank Derek Jensen for helpful discussions. This project was supported by grants from the National Institute of General Medical Sciences (P20GM103440), the American Cancer Society (IRG-103719), and a UNLV Faculty Opportunity Award.

Supplementary material

10534_2014_9783_MOESM1_ESM.pdf (177 kb)
Supplementary material 1 (PDF 177 kb)


  1. Boehme KA, Kulikov R, Blattner C (2008) p53 stabilization in response to DNA damage requires Akt/PKB and DNA-PK. Proc Natl Acad Sci USA 105:7785–7790PubMedCentralPubMedCrossRefGoogle Scholar
  2. Cheng K, Creacy S, Larner J (1983) ‘Insulin-like’ effects of lithium ion on isolated rat adipocytes. II. Specific activation of glycogen synthase. Mol Cell Biochem 56:183–189PubMedGoogle Scholar
  3. Coates SS, Lehnert BE, Sharma S, Kindell SM, Gary RK (2007) Beryllium induces premature senescence in human fibroblasts. J Pharmacol Exp Ther 322:70–79PubMedCrossRefGoogle Scholar
  4. Cohen P, Frame S (2001) The renaissance of GSK3. Nat Rev Mol Cell Biol 2:769–776PubMedCrossRefGoogle Scholar
  5. Cole AR (2013) Glycogen synthase kinase 3 substrates in mood disorders and schizophrenia. FEBS J 280:5213–5227PubMedCrossRefGoogle Scholar
  6. Cole A, Frame S, Cohen P (2004) Further evidence that the tyrosine phosphorylation of glycogen synthase kinase-3 (GSK3) in mammalian cells is an autophosphorylation event. Biochem J 377:249–255PubMedCentralPubMedCrossRefGoogle Scholar
  7. Ding VW, Chen RH, McCormick F (2000) Differential regulation of glycogen synthase kinase 3β by insulin and Wnt signaling. J Biol Chem 275:32475–32481PubMedCrossRefGoogle Scholar
  8. Doble BW, Woodgett JR (2003) GSK-3: tricks of the trade for a multi-tasking kinase. J Cell Sci 116:1175–1186PubMedCentralPubMedCrossRefGoogle Scholar
  9. Fang X, Yu SX, Lu Y, Bast RC Jr, Woodgett JR, Mills GB (2000) Phosphorylation and inactivation of glycogen synthase kinase 3 by protein kinase A. Proc Natl Acad Sci USA 97:11960–11965PubMedCentralPubMedCrossRefGoogle Scholar
  10. Force T, Woodgett JR (2009) Unique and overlapping functions of GSK-3 isoforms in cell differentiation and proliferation and cardiovascular development. J Biol Chem 284:9643–9647PubMedCentralPubMedCrossRefGoogle Scholar
  11. Gorjala P, Gary RK (2010) Beryllium sulfate induces p21CDKN1A expression and a senescence-like cell cycle arrest in susceptible cancer cell types. Biometals 23:1061–1073PubMedCentralPubMedCrossRefGoogle Scholar
  12. Horn RS, Walaas O, Walaas E (1973) The influence of sodium, potassium and lithium on the response of glycogen synthetase I to insulin and epinephrine in the isolated rat diaphragm. Biochim Biophys Acta 313:296–309PubMedCrossRefGoogle Scholar
  13. Jope RS (2003) Lithium and GSK-3: one inhibitor, two inhibitory actions, multiple outcomes. Trends Pharmacol Sci 24:441–443PubMedCrossRefGoogle Scholar
  14. Jope RS, Johnson GV (2003) The glamour and gloom of glycogen synthase kinase-3. Trends Biochem Sci 29:95–102CrossRefGoogle Scholar
  15. Klein PS, Melton DA (1996) A molecular mechanism for the effect of lithium on development. Proc Natl Acad Sci USA 93:8455–8459PubMedCentralPubMedCrossRefGoogle Scholar
  16. Kulikov R, Boehme KA, Blattner C (2005) Glycogen synthase kinase 3-dependent phosphorylation of Mdm2 regulates p53 abundance. Mol Cell Biol 25:7170–7180PubMedCentralPubMedCrossRefGoogle Scholar
  17. Lehnert NM, Gary RK, Marrone BL, Lehnert BE (2001) Inhibition of normal human lung fibroblast growth by beryllium. Toxicology 160:119–127PubMedCrossRefGoogle Scholar
  18. Li L, Kozlowski K, Wegner B, Rashid T, Yeung T, Holmes C, Ballermann BJ (2007) Phosphorylation of TIMAP by glycogen synthase kinase-3beta activates its associated protein phosphatase 1. J Biol Chem 282:25960–25969PubMedCrossRefGoogle Scholar
  19. Liu F, Liang Z, Shi J, Yin D, El-Akkad E, Grundke-Iqbal I, Iqbal K, Gong CX (2006) PKA modulates GSK-3beta- and cdk5-catalyzed phosphorylation of tau in site- and kinase-specific manners. FEBS Lett 580:6269–6274PubMedCentralPubMedCrossRefGoogle Scholar
  20. Mao CD, Hoang P, DiCorleto PE (2001) Lithium inhibits cell cycle progression and induces stabilization of p53 in bovine aortic endothelial cells. J Biol Chem 276:26180–26188PubMedCrossRefGoogle Scholar
  21. Maurer U, Preiss F, Brauns-Schubert P, Schlicher L, Charvet C (2014) GSK-3—at the crossroads of cell death and survival. J Cell Sci 127:1369–1378PubMedCrossRefGoogle Scholar
  22. Niles AL, Moravec RA, Eric Hesselberth P, Scurria MA, Daily WJ, Riss TL (2007) A homogeneous assay to measure live and dead cells in the same sample by detecting different protease markers. Anal Biochem 366:197–206PubMedCrossRefGoogle Scholar
  23. Olsher U, Izatt RM, Bradshaw JS, Dalley NK (1991) Coordination chemistry of lithium ion: a crystal and molecular structure review. Chem Rev 91:137–164CrossRefGoogle Scholar
  24. Pittet PA, Elbaze G, Helm L, Merbach AE (1990) Tetrasolventoberyllium(II): high-pressure evidence for a sterically controlled solvent-exchange-mechanism crossover. Inorg Chem 29:1936–1942CrossRefGoogle Scholar
  25. Ryves WJ, Harwood AJ (2001) Lithium inhibits glycogen synthase kinase-3 by competition for magnesium. Biochem Biophys Res Commun 280:720–725PubMedCrossRefGoogle Scholar
  26. Ryves WJ, Dajani R, Pearl L, Harwood AJ (2002) Glycogen synthase kinase-3 inhibition by lithium and beryllium suggests the presence of two magnesium binding sites. Biochem Biophys Res Commun 290:967–972PubMedCrossRefGoogle Scholar
  27. Seo YH, Jung HJ, Shin HT, Kim YM, Yim H, Chung HY, Lim IK, Yoon G (2008) Enhanced glycogenesis is involved in cellular senescence via GSK3/GS modulation. Aging Cell 7:894–907PubMedCrossRefGoogle Scholar
  28. Sheridan CM, Heist EK, Beals CR, Crabtree GR, Gardner P (2002) Protein kinase A negatively modulates the nuclear accumulation of NF-ATc1 by priming for subsequent phosphorylation by glycogen synthase kinase-3. J Biol Chem 277:48664–48676PubMedCrossRefGoogle Scholar
  29. Stambolic V, Ruel L, Woodgett JR (1996) Lithium inhibits glycogen synthase kinase-3 activity and mimics wingless signalling in intact cells. Curr Biol 6:1664–1668PubMedCrossRefGoogle Scholar
  30. Sutherland C, Leighton IA, Cohen P (1993) Inactivation of glycogen synthase kinase-3 beta by phosphorylation: new kinase connections in insulin and growth-factor signalling. Biochem J 296:15–19PubMedCentralPubMedGoogle Scholar
  31. Tanji C, Yamamoto H, Yorioka N, Kohno N, Kikuchi K, Kikuchi A (2002) A-kinase anchoring protein AKAP220 binds to glycogen synthase kinase-3beta (GSK-3beta) and mediates protein kinase A-dependent inhibition of GSK-3beta. J Biol Chem 277:36955–36961PubMedCrossRefGoogle Scholar
  32. Turenne GA, Price BD (2001) Glycogen synthase kinase3 beta phosphorylates serine 33 of p53 and activates p53’s transcriptional activity. BMC Cell Biol 2:12. doi: 10.1186/1471-2121-2-12 PubMedCentralPubMedCrossRefGoogle Scholar
  33. Watcharasit P, Bijur GN, Zmijewski JW, Song L, Zmijewska A, Chen X, Johnson GV, Jope RS (2002) Direct, activating interaction between glycogen synthase kinase-3beta and p53 after DNA damage. Proc Natl Acad Sci USA 99:7951–7955PubMedCentralPubMedCrossRefGoogle Scholar
  34. Watcharasit P, Bijur GN, Song L, Zhu J, Chen X, Jope RS (2003) Glycogen synthase kinase-3beta (GSK3beta) binds to and promotes the actions of p53. J Biol Chem 278:48872–48879PubMedCentralPubMedCrossRefGoogle Scholar
  35. Ye X, Zerlanko B, Kennedy A, Banumathy G, Zhang R, Adams PD (2007) Downregulation of Wnt signaling is a trigger for formation of facultative heterochromatin and onset of cell senescence in primary human cells. Mol Cell 27:183–196PubMedCentralPubMedCrossRefGoogle Scholar
  36. Zhang F, Phiel CJ, Spece L, Gurvich N, Klein PS (2003) Inhibitory phosphorylation of glycogen synthase kinase-3 (GSK-3) in response to lithium: evidence for autoregulation of GSK-3. J Biol Chem 278:33067–33077PubMedCrossRefGoogle Scholar
  37. Zmijewski JW, Jope RS (2004) Nuclear accumulation of glycogen synthase kinase-3 during replicative senescence of human fibroblasts. Aging Cell 3:309–317PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Swapna R. Mudireddy
    • 1
  • Ataur Rahman Mohammed Abdul
    • 1
  • Priyatham Gorjala
    • 1
  • Ronald K. Gary
    • 1
  1. 1.Department of ChemistryUniversity of Nevada, Las VegasLas VegasUSA

Personalised recommendations