Caspase-3 mediated release of SAC domain containing fragment from Par-4 is necessary for the sphingosine-induced apoptosis in Jurkat cells

Journal of Molecular Signaling

Sehamuddin Galadari¹  Mahendra Patel¹  Shahanas Chathoth¹  Jaleel Kizhakkayil¹  Anees Rahman¹  Siraj Pallichankandy¹  Faisal Thayyullathil¹ 

[1] Cell Signaling Laboratory, Department of Biochemistry, College of Medicine and Health Sciences, UAE University, P.O. Box 17666, Al Ain, Abu Dhabi, UAE

关键词: Sphingosine;    Cleavage;    SAC domain;    Apoptosis;    Caspase;    Par-4;   

Others  :  802736
DOI  :  10.1186/1750-2187-8-2

 received in 2012-09-24, accepted in 2013-02-23,  发布年份 2013

【 摘 要 】

Background

Prostate apoptosis response-4 (Par-4) is a tumor-suppressor protein that selectively activates and induces apoptosis in cancer cells, but not in normal cells. The cancer specific pro-apoptotic function of Par-4 is encoded in its centrally located SAC (Selective for Apoptosis induction in Cancer cells) domain (amino acids 137–195). The SAC domain itself is capable of nuclear entry, caspase activation, inhibition of NF-κB activity, and induction of apoptosis in cancer cells. However, the precise mechanism(s) of how the SAC domain is released from Par-4, in response to apoptotic stimulation, is not well explored.

Results

In this study, we demonstrate for the first time that sphingosine (SPH), a member of the sphingolipid family, induces caspase-dependant cleavage of Par-4, leading to the release of SAC domain containing fragment from it. Par-4 is cleaved at the EEPD131G site on incubation with caspase-3 in vitro, and by treating cells with several anti-cancer agents. The caspase-3 mediated cleavage of Par-4 is blocked by addition of the pan-caspase inhibitor z-VAD-fmk, caspase-3 specific inhibitor Ac-DEVD-CHO, and by introduction of alanine substitution for D131 residue. Moreover, suppression of SPH-induced Akt dephosphorylation also abrogated the caspase dependant cleavage of Par-4.

Conclusion

Evidence provided here shows that Par-4 is cleaved by caspase-3 during SPH-induced apoptosis. Cleavage of Par-4 leads to the generation of SAC domain containing fragment which may possibly be essential and sufficient to induce or augment apoptosis in cancer cells.

【 授权许可】

   
2013 Thayyullathil et al; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Rllis RE, Yuan JY, Horvitz HR: Mechanisms and functions of cell death. Annu Rev Cell Biol 1991, 7:663-698.
• [2]Bauer J, Wekerle H, Lassmann H: Apoptosis in brain-specific autoimmune disease. Curr Opin Immunol 1995, 7:839-843.
• [3]Crawford ED, Wells JA: Caspase substrates and cellular remodeling. Annu Rev Biochem 2011, 80:1055-1087.
• [4]Earnshaw WC, Martins LM, Kaufmann SH: Mammalian caspases: structure, activation, substrates, and functions during apoptosis. Annu Rev Biochem 1999, 68:383-424.
• [5]Sayan BS, Sayan AE, Knight RA, Melino G, Cohen GM: p53 is cleaved by caspases generating fragments localizing to mitochondria. J Biol Chem 2006, 281:13566-13573.
• [6]Sells SF, Wood DP Jr, Joshi-Barve SS, Muthukumar S, Jacob RJ, Crist SA, Humphreys S, Rangnekar VM: Commonality of the gene programs induced by effectors of apoptosis in androgen-dependent and -independent prostate cells. Cell Growth Differ 1994, 5:457-466.
• [7]Sells SF, Han SS, Muthukkumar S, Maddiwar N, Johnstone R, Boghaert E, Gillis D, Liu G, Nair P, Monnig S, Collini P, Mattson MP, Sukhatme VP, Zimmer SG, Wood DP Jr, McRoberts JW, Shi Y, Rangnekar VM: Expression and function of the leucine zipper protein Par-4 in apoptosis. Mol Cell Biol 1997, 17:3823-3832.
• [8]El-Guendy N, Zhao Y, Gurumurthy S, Burikhanov R, Rangnekar VM: Identification of a unique core domain of par-4 sufficient for selective apoptosis induction in cancer cells. Mol Cell Biol 2003, 23:5516-5525.
• [9]Ohta H, Sweeney EA, Masamune A, Yatomi Y, Hakomori S, Igarashi Y: Induction of apoptosis by sphingosine in human leukemic HL-60 cells: a possible endogenous modulator of apoptotic DNA fragmentation occurring during phorbol ester-induced differentiation. Cancer Res 1995, 55:691-697.
• [10]Jarvis WD, Fornari FA, Traylor RS, Martin HA, Kramer LB, Erukulla RK, Bittman R, Grant S: Induction of apoptosis and potentiation of ceramide-mediated cytotoxicity by sphingoid bases in human myeloid leukemia cells. J Biol Chem 1996, 271:8275-8284.
• [11]Cuvillier O, Edsall L, Spiegel S: Involvement of sphingosine in mitochondria-dependent Fas-induced apoptosis of type II Jurkat T cells. J Biol Chem 2000, 275:15691-15700.
• [12]Thayyullathil F, Chathoth S, Shahin A, Kizhakkayil J, Hago A, Patel M, Galadari S: Protein phosphatase 1-dependent dephosphorylation of Akt is the prime signaling event in sphingosine-induced apoptosis in Jurkat cells. J Cell Biochem 2011, 112:1138-1153.
• [13]Phillips DC, Martin S, Doyle BT, Houghton JA: Sphingosine-induced apoptosis in rhabdomyosarcoma cell lines is dependent on pre-mitochondrial Bax activation and post-mitochondrial caspases. Cancer Res 2007, 67:756-764.
• [14]Cuvillier O: Sphingosine in apoptosis signaling. Biochim Biophys Acta 2002, 1585:153-162.
• [15]Hung WC, Chang HC, Chuang LY: Activation of caspase-3-like proteases in apoptosis induced by sphingosine and other long-chain bases in Hep3B hepatoma cells. Biochem J 1999, 338:161-166.
• [16]Dudek H, Datta SR, Franke TF, Birnbaum MJ, Yao R, Cooper GM, Segal RA, Kaplan DR, Greenberg ME: Regulation of neuronal survival by the serine-threonine protein kinase Akt. Science 1997, 275:661-665.
• [17]Alessi DR, Andjelkovic M, Caudwell B, Cron P, Morrice N, Cohen P, Hemmings BA: Mechanism of activation of protein kinase B by insulin and IGF-1. EMBO J 1996, 15:6541-6551.
• [18]Alessi DR, James SR, Downes CP, Holmes AB, Gaffney PR, Reese CB, Cohen P: Characterization of a 3-phosphoinositide-dependent protein kinase which phosphorylates and activates protein kinase Balpha. Curr Biol 1997, 7:261-269.
• [19]Goswami A, Burikhanov R, de Thonel A, Fujita N, Goswami M, Zhao Y, Eriksson JE, Tsuruo T, Rangnekar VM: Binding and phosphorylation of par-4 by akt is essential for cancer cell survival. Mol Cell 2005, 20:33-44.
• [20]Goswami A, Ranganathan P, Rangnekar VM: The phosphoinositide 3-kinase/Akt1/Par-4 axis: a cancer-selective therapeutic target. Cancer Res 2006, 66:2889-2892.
• [21]Cross DA, Alessi DR, Cohen P, Andjelkovich M, Hemmings BA: Inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B. Nature 1995, 378:785-789.
• [22]del Peso L, Gonzalez-Garcia M, Page C, Herrera R, Nunez G: Interleukin-3-induced phosphorylation of BAD through the protein kinase Akt. Science 1997, 278:687-689.
• [23]Cardone MH, Roy N, Stennicke HR, Salvesen GS, Franke TF, Stanbridge E, Frisch S, Reed JC: Regulation of cell death protease caspase-9 by phosphorylation. Science 1998, 282:1318-1321.
• [24]Dan HC, Sun M, Kaneko S, Feldman RI, Nicosia SV, Wang HG, Tsang BK, Cheng JQ: Akt phosphorylation and stabilization of X-linked inhibitor of apoptosis protein (XIAP). J Biol Chem 2004, 279:5405-5412.
• [25]Kim AH, Khursigara G, Sun X, Franke TF, Chao MV: Akt phosphorylates and negatively regulates apoptosis signal-regulating kinase 1. Mol Cell Biol 2001, 21:893-901.
• [26]Zhou BP, Liao Y, Xia W, Zou Y, Spohn B, Hung MC: HER-2/neu induces p53 ubiquitination via Akt-mediated MDM2 phosphorylation. Nat Cell Biol 2001, 3:973-982.
• [27]Chen CS, Weng SC, Tseng PH, Lin HP, Chen CS: Histone acetylation-independent effect of histone deacetylase inhibitors on Akt through the reshuffling of protein phosphatase 1 complexes. J Biol Chem 2005, 280:38879-38887.
• [28]Meier R, Thelen M, Hemmings BA: Inactivation and dephosphorylation of protein kinase Balpha (PKBalpha) promoted by hyperosmotic stress. EMBO J 1998, 17:7294-7303.
• [29]Widmann C, Gibson S, Johnson GL: Caspase-dependent cleavage of signaling proteins during apoptosis. A turn-off mechanism for anti-apoptotic signals. J Biol Chem 1998, 273:7141-7147.
• [30]Chaudhry P, Singh M, Parent S, Asselin E: Prostate apoptosis response 4 (Par-4), a novel substrate of caspase-3 during apoptosis activation. Mol Cell Biol 2012, 32:826-839.
• [31]Merrill AH Jr, van Echten G, Wang E, Sandhoff K: Fumonisin B1 inhibits sphingosine (sphinganine) N-acyltransferase and de novo sphingolipid biosynthesis in cultured neurons in situ. J Biol Chem 1993, 268:27299-27306.
• [32]Fischer U, Janicke RU, Schulze-Osthoff K: Many cuts to ruin: a comprehensive update of caspase substrates. Cell Death Differ 2003, 10:76-100.
• [33]Janicke RU, Sprengart ML, Wati MR, Porter AG: Caspase-3 is required for DNA fragmentation and morphological changes associated with apoptosis. J Biol Chem 1998, 273:9357-9360.
• [34]Janicke RU, Ng P, Sprengart ML, Porter AG: Caspase-3 is required for alpha-fodrin cleavage but dispensable for cleavage of other death substrates in apoptosis. J Biol Chem 1998, 273:15540-15545.
• [35]Mooney LM, Al-Sakkaf KA, Brown BL, Dobson PR: Apoptotic mechanisms in T47D and MCF-7 human breast cancer cells. Br J Cancer 2002, 87:909-917.
• [36]Fattman CL, Delach SM, Dou QP, Johnson DE: Sequential two-step cleavage of the retinoblastoma protein by caspase-3/-7 during etoposide-induced apoptosis. Oncogene 2001, 20:2918-2926.
• [37]Slee EA, Adrain C, Martin SJ: Executioner caspase-3, -6, and −7 perform distinct, non-redundant roles during the demolition phase of apoptosis. J Biol Chem 2001, 276:7320-7326.
• [38]Talanian RV, Quinlan C, Trautz S, Hackett MC, Mankovich JA, Banach D, Ghayur T, Brady KD, Wong WW: Substrate specificities of caspase family proteases. J Biol Chem 1997, 272:9677-9682.
• [39]Thornberry NA, Rano TA, Peterson EP, Rasper DM, Timkey T, Garcia-Calvo M, Houtzager VM, Nordstrom PA, Roy S, Vaillancourt JP, Chapman KT, Nicholson DW: Combinatorial approach defines specificities of members of the caspase family and granzyme B. Functional relationships established for key mediators of apoptosis. J Biol Chem 1997, 272:17907-17911.
• [40]Bae SS, Perry DK, Oh YS, Choi JH, Galadari S, Ghayur T, Ryu SH, Hannun YA, Suh PG: Proteolytic cleavage of phospholipase C-gamma1 during apoptosis in Molt-4 cells. FASEB J 2000, 2000(14):1083-1092.
• [41]Samejima K, Svingen PA, Basi GS, Kottke T, Mesner PW Jr, Stewart L, Durrieu F, Poirier GG, Alnemri ES, Champoux JJ, Kaufmann SH, Earnshaw WC: Caspase-mediated cleavage of DNA topoisomerase I at unconventional sites during apoptosis. J Biol Chem 1999, 274:4335-4340.
• [42]Gurumurthy S, Goswami A, Vasudevan KM, Rangnekar VM: Phosphorylation of Par-4 by protein kinase A is critical for apoptosis. Mol Cell Biol 2005, 25:1146-1161.
• [43]Thayyullathil F, Chathoth S, Kizhakkayil J, Galadari A, Hago A, Patel M, Galadari S: Glutathione selectively inhibits Doxorubicin induced phosphorylation of p53Ser15, caspase dependent ceramide production and apoptosis in human leukemic cells. Biochem Biophys Res Commun 2011, 411:1-6.