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Diabetologia

Erschienen in:

01.04.2013 | Article

14-3-3 proteins are essential signalling hubs for beta cell survival

verfasst von: G. E. Lim, M. Piske, J. D. Johnson

Erschienen in: Diabetologia | Ausgabe 4/2013

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Abstract

Aims/hypothesis

Diabetes is characterised by pancreatic beta cell death and dysfunction, resulting from unbalanced pro-survival and pro-death signalling. The 14-3-3 proteins are molecular adaptors that integrate numerous signalling pathways, including the v-raf-leukaemia viral oncogene 1 (RAF1)/B cell leukaemia/lymphoma 2 (BCL-2)-associated agonist of cell death (BAD) pathway, which we have previously implicated in insulin-dependent beta cell survival. Nevertheless, the roles of 14-3-3 proteins in beta cell fate and function have not been investigated.

Methods

We examined the abundance, localisation, modulation and roles of 14-3-3 proteins using quantitative RT-PCR, immunoblot or imaging. MIN6 cells or mouse islets cells were manipulated with inhibitors, short interfering RNA (siRNA) or plasmids overexpressing 14-3-3.

Results

We first characterised the abundance and subcellular location of all seven 14-3-3 isoforms in mouse and human beta cells. Most isoforms were cytoplasmic, except 14-3-3σ, which appeared to be nuclear. Analysis of 14-3-3 abundance under stress conditions revealed distinct modulation in mouse islets and MIN6 cells. Generalised 14-3-3 inhibition promoted apoptosis and dysfunction, and siRNA-mediated knockdown revealed isoform-specific roles in caspase-3-dependent beta cell apoptosis, with a clear role for 14-3-3ζ. Overabundance of 14-3-3ζ sequestered BAD–BCL2-associated X protein (BAX) from mitochondria, attenuated Dp5 (also known as Hrk) and Puma (also known as Bbc3) induction, and increased survival in response to pro-inflammatory cytokines or thapsigargin. Anti-apoptotic insulin treatment increased the sequestration of BAD/BAX by 14-3-3ζ. BAD mutants that were unable to bind 14-3-3ζ localised to mitochondria and induced apoptosis.

Conclusions/interpretation

This first study of the 14-3-3 family in beta cells revealed specific regulation, localisation and anti-apoptotic roles among the isoforms. Focus on 14-3-3ζ revealed its importance in preventing BAD–BAX mitochondrial localisation and protecting beta cells from multiple stresses. Thus, some 14-3-3 proteins are pro-survival signalling hubs.

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Prentki M, Nolan CJ (2006) Islet beta cell failure in type 2 diabetes. J Clin Invest 116:1802–1812PubMedCrossRef

Harlan DM, Kenyon NS, Korsgren O, Roep BO (2009) Current advances and travails in islet transplantation. Diabetes 58:2175–2184PubMedCrossRef

Scott JD, Pawson T (2009) Cell signaling in space and time: where proteins come together and when they're apart. Science 326:1220–1224PubMedCrossRef

Morrison DK (2009) The 14-3-3 proteins: integrators of diverse signaling cues that impact cell fate and cancer development. Trends Cell Biol 19:16–23PubMedCrossRef

Petosa C, Masters SC, Bankston LA et al (1998) 14-3-3zeta binds a phosphorylated Raf peptide and an unphosphorylated peptide via its conserved amphipathic groove. J Biol Chem 273:16305–16310PubMedCrossRef

Rosenquist M, Sehnke P, Ferl RJ, Sommarin M, Larsson C (2000) Evolution of the 14-3-3 protein family: does the large number of isoforms in multicellular organisms reflect functional specificity? J Mol Evol 51:446–458PubMed

Subramanian RR, Masters SC, Zhang H, Fu H (2001) Functional conservation of 14-3-3 isoforms in inhibiting bad-induced apoptosis. Exp Cell Res 271:142–151PubMedCrossRef

Lau JM, Wu C, Muslin AJ (2006) Differential role of 14-3-3 family members in Xenopus development. Dev Dyn 235:1761–1776PubMedCrossRef

Xing H, Zhang S, Weinheimer C, Kovacs A, Muslin AJ (2000) 14-3-3 proteins block apoptosis and differentially regulate MAPK cascades. EMBO J 19:349–358PubMedCrossRef

10.

Datta SR, Katsov A, Hu L et al (2000) 14-3-3 proteins and survival kinases cooperate to inactivate BAD by BH3 domain phosphorylation. Mol Cell 6:41–51PubMed

11.

Masters SC, Fu H (2001) 14-3-3 proteins mediate an essential anti-apoptotic signal. J Biol Chem 276:45193–45200PubMedCrossRef

12.

Chiang CW, Kanies C, Kim KW et al (2003) Protein phosphatase 2A dephosphorylation of phosphoserine 112 plays the gatekeeper role for BAD-mediated apoptosis. Mol Cell Biol 23:6350–6362PubMedCrossRef

13.

Fantl WJ, Muslin AJ, Kikuchi A et al (1994) Activation of Raf-1 by 14-3-3 proteins. Nature 371:612–614PubMedCrossRef

14.

Alejandro EU, Johnson JD (2008) Inhibition of Raf-1 alters multiple downstream pathways to induce pancreatic beta-cell apoptosis. J Biol Chem 283:2407–2417PubMedCrossRef

15.

Alejandro EU, Kalynyak TB, Taghizadeh F et al (2010) Acute insulin signaling in pancreatic beta-cells is mediated by multiple Raf-1 dependent pathways. Endocrinology 151:502–512PubMedCrossRef

16.

Alejandro EU, Lim GE, Mehran AE et al (2011) Pancreatic beta-cell Raf-1 is required for glucose tolerance, insulin secretion, and insulin 2 transcription. FASEB J 25:3884–3895PubMedCrossRef

17.

Beith JL, Alejandro EU, Johnson JD (2008) Insulin stimulates primary beta-cell proliferation via Raf-1 kinase. Endocrinology 149:2251–2260PubMedCrossRef

18.

Barry EF, Felquer FA, Powell JA et al (2009) 14-3-3:Shc scaffolds integrate phosphoserine and phosphotyrosine signaling to regulate phosphatidylinositol 3-kinase activation and cell survival. J Biol Chem 284:12080–12090PubMedCrossRef

19.

Szabat M, Luciani DS, Piret JM, Johnson JD (2009) Maturation of adult beta-cells revealed using a Pdx1/insulin dual-reporter lentivirus. Endocrinology 150:1627–1635PubMedCrossRef

20.

Szabat M, Pourghaderi P, Soukhatcheva G et al (2011) Kinetics and genomic profiling of adult human and mouse beta-cell maturation. Islets 3:175–187PubMedCrossRef

21.

He M, Zhang J, Shao L et al (2006) Upregulation of 14-3-3 isoforms in acute rat myocardial injuries induced by burn and lipopolysaccharide. Clin Exp Pharmacol Physiol 33:374–380PubMedCrossRef

22.

Kuzelova K, Grebenova D, Pluskalova M, Kavan D, Halada P, Hrkal Z (2009) Isoform-specific cleavage of 14-3-3 proteins in apoptotic JURL-MK1 cells. J Cell Biochem 106:673–681PubMedCrossRef

23.

Akerfeldt MC, Howes J, Chan JY et al (2008) Cytokine-induced beta-cell death is independent of endoplasmic reticulum stress signaling. Diabetes 57:3034–3044PubMedCrossRef

24.

Lai E, Bikopoulos G, Wheeler MB, Rozakis-Adcock M, Volchuk A (2008) Differential activation of ER stress and apoptosis in response to chronically elevated free fatty acids in pancreatic beta-cells. Am J Phyisol Endocrinol Metabol 294:E540–E550CrossRef

25.

Masters SC, Yang H, Datta SR, Greenberg ME, Fu H (2001) 14-3-3 inhibits Bad-induced cell death through interaction with serine-136. Mol Pharmacol 60:1325–1331PubMed

26.

Tsuruta F, Sunayama J, Mori Y et al (2004) JNK promotes Bax translocation to mitochondria through phosphorylation of 14-3-3 proteins. EMBO J 23:1889–1899PubMedCrossRef

27.

Morgan A, Burgoyne RD (1992) Exo1 and Exo2 proteins stimulate calcium-dependent exocytosis in permeabilized adrenal chromaffin cells. Nature 355:833–836PubMedCrossRef

28.

Johnson JD, Alejandro EU (2008) Control of pancreatic beta-cell fate by insulin signaling: the sweet spot hypothesis. Cell Cycle 7:1343–1347PubMedCrossRef

29.

Cnop M, Welsh N, Jonas JC, Jorns A, Lenzen S, Eizirik DL (2005) Mechanisms of pancreatic beta-cell death in type 1 and type 2 diabetes: many differences, few similarities. Diabetes 54(Suppl 2):S97–S107PubMedCrossRef

30.

Thomas HE, Biden TJ (2009) Bad news for beta-cell apoptosis. Diabetes 58:1725–1727PubMedCrossRef

31.

Grunnet LG, Aikin R, Tonnesen MF et al (2009) Proinflammatory cytokines activate the intrinsic apoptotic pathway in beta-cells. Diabetes 58:1807–1815PubMedCrossRef

32.

Polzien L, Baljuls A, Rennefahrt UE et al (2009) Identification of novel in vivo phosphorylation sites of the human proapoptotic protein BAD: pore-forming activity of BAD is regulated by phosphorylation. J Biol Chem 284:28004–28020PubMedCrossRef

33.

Johnson JD, Bernal-Mizrachi E, Alejandro EU et al (2006) Insulin protects islets from apoptosis via Pdx1 and specific changes in the human islet proteome. Proc Natl Acad Sci U S A 103:19575–19580PubMedCrossRef

34.

Virdee K, Parone PA, Tolkovsky AM (2000) Phosphorylation of the pro-apoptotic protein BAD on serine 155, a novel site, contributes to cell survival. Current Biol: CB 10:1151–1154CrossRef

35.

Zhou J, Shao Z, Kerkela R et al (2009) Serine 58 of 14-3-3zeta is a molecular switch regulating ASK1 and oxidant stress-induced cell death. Mol Cell Biol 29:4167–4176PubMedCrossRef

36.

Aguilera C, Fernandez-Majada V, Ingles-Esteve J, Rodilla V, Bigas A, Espinosa L (2006) Efficient nuclear export of p65-IkappaBalpha complexes requires 14-3-3 proteins. J Cell Sci 119:3695–3704PubMedCrossRef

37.

Barthson J, Germano CM, Moore F et al (2011) Cytokines tumor necrosis factor-alpha and interferon-gamma induce pancreatic beta-cell apoptosis through STAT1-mediated Bim protein activation. J Biol Chem 286:39632–39643PubMedCrossRef

38.

Gurzov EN, Germano CM, Cunha DA et al (2010) p53 up-regulated modulator of apoptosis (PUMA) activation contributes to pancreatic beta-cell apoptosis induced by proinflammatory cytokines and endoplasmic reticulum stress. J Biol Chem 285:19910–19920PubMedCrossRef

39.

Gurzov EN, Ortis F, Cunha DA et al (2009) Signaling by IL-1beta + IFN-gamma and ER stress converge on DP5/Hrk activation: a novel mechanism for pancreatic beta-cell apoptosis. Cell Death Diff 16:1539–1550CrossRef

40.

Wang P, Qiu W, Dudgeon C et al (2009) PUMA is directly activated by NF-kappaB and contributes to TNF-alpha-induced apoptosis. Cell Death Diff 16:1192–1202CrossRef

41.

Quoyer J, Longuet C, Broca C et al (2009) GLP-1 mediates antiapoptotic effect by phosphorylating Bad through a beta-arrestin 1-mediated ERK1/2 activation in pancreatic beta-cells. J Biol Chem 285:1989–2002PubMedCrossRef

42.

Gardino AK, Smerdon SJ, Yaffe MB (2006) Structural determinants of 14-3-3 binding specificities and regulation of subcellular localization of 14-3-3-ligand complexes: a comparison of the X-ray crystal structures of all human 14-3-3 isoforms. Semin Canc Biol 16:173–182CrossRef

43.

Ewing RM, Chu P, Elisma F et al (2007) Large-scale mapping of human protein-protein interactions by mass spectrometry. Mol Syst Biol 3:89PubMedCrossRef

44.

Zha J, Harada H, Yang E, Jockel J, Korsmeyer SJ (1996) Serine phosphorylation of death agonist BAD in response to survival factor results in binding to 14-3-3 not BCL-X(L). Cell 87:619–628PubMedCrossRef

45.

Pozuelo-Rubio M (2010) Proteomic and biochemical analysis of 14-3-3-binding proteins during C2-ceramide-induced apoptosis. FEBS J 277:3321–3342PubMedCrossRef

46.

Luciani DS, White SA, Widenmaier SB et al (2012) Bcl-2 and Bcl-xL suppress glucose signaling in pancreatic beta-cells. Diabetes. doi:10.2337/db11-1464

47.

Danial NN, Walensky LD, Zhang CY et al (2008) Dual role of proapoptotic BAD in insulin secretion and beta cell survival. Nat Med 14:144–153PubMedCrossRef

48.

Zhao J, Meyerkord CL, Du Y, Khuri FR, Fu H (2011) 14-3-3 proteins as potential therapeutic targets. Semin Cell Dev Biol 22:705–712PubMedCrossRef

Titel: 14-3-3 proteins are essential signalling hubs for beta cell survival
verfasst von: G. E. Lim
M. Piske
J. D. Johnson
Publikationsdatum: 01.04.2013
Verlag: Springer-Verlag
Erschienen in: Diabetologia / Ausgabe 4/2013
Print ISSN: 0012-186X
Elektronische ISSN: 1432-0428
DOI: https://doi.org/10.1007/s00125-012-2820-x

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