Abstract
We have recently reported that EGF triggers an original form of cell death in pituitary cell line (GH4C1) with a phenotype sharing some characteristics of both apoptosis (internucleosomal DNA fragmentation) and paraptosis (caspase-independence and cytoplasmic vacuolization). However, the endonuclease involved in EGF-induced DNA fragmentation has not been assessed so far. In the present work we therefore further explored the putative paraptosis involvement in EGF-induced cell death and asked whether L-DNaseII might be involved. Indeed, this endonuclease is known to mediate internucleosomal DNA fragmentation in caspase independent manner. Our Western blot, immunocytochemistry and enzymatic measurement assays show that EGF triggers a cleavage of Leukocyte Elastase Inhibitor (LEI) precursor into L-DNaseII, its subsequent enzymatic activation and nuclear translocation thus pointing to the involvement of this endonuclease pathway in caspase-independent DNA fragmentation. In addition, EGF-induced cell death can be blocked by paraptosis inhibitor AIP-1/Alix, but not with its anti-apoptotic C-terminal fragment (Alix-CT). Altogether these data suggest that EGF-induced cell death defines a novel, L-DNaseII-mediated form of paraptosis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Clarke P. Developmental cell death: Morphological diversity and multiple mechanisms. Anat Embryol 1990; 181: 195–213.
Budihardjo I, Oliver H, Lutter M, Luo X, Wang X. Biochemical pathways of caspase activation during apoptosis. Annu Rev Cell Dev Biol 1999; 15: 269–290.
Jaattela M, Tschopp J. Caspase-independent cell death in T lymphocytes. Nature Immunol 2003; 4: 416–423.
Nagata S. Apoptosis by death factor. Cell. 1997; 88: 355–365.
Rao R, Hermel E, Castro-Obregon S, et al. Coupling Endoplasmic Reticulum stress to the cell death program (Mechanism of caspase activation). J Biol Chem 2001; 276: 33869–33874.
Forcet C, Ye X, Granger L. et al. The dependence receptor DCC (deleted in colorectal cancer) defines an alternative mechanism for caspase activation. Proc Natl Acad Sci 2001; 98: 3416–3421.
Chi S, Kitanaka C, Noguchi K, et al. Oncogenic Ras triggers cell suicide through the activation of a caspase-independent cell death program in human cancer cells. Oncogene 1999; 18: 2281–2290.
Sperandio S, de Belle I, Bredesen D. An alternative, nonapoptotic form of programmed cell death. Proc Natl Acad Sci 2000; 97: 14376–14381.
Castro-Obregon S, del Rio G, Chen S, et al. A ligand-receptor pair that triggers a non-apoptotic cell death. Cell Death Differ 2002; 9: 807–819.
Sperandio S, Poksay K, de Belle I, et al. Paraptosis: Mediation by MAP kinases and inhibition by AIP-1/Alix. Cell Death Differ 2004; 11: 1066–1075.
Fombonne J, Reix S, Rasolonjanahary S, et al. EGF triggers an original caspase-independent pituitary cell death with heterogenous phenotype. Mol Biol Cell 2004; 15: 4938–4948.
Evans C, Aguilera R. DNaseII: Genes, enzymes and functions. Gene 2003; 322: 1–15.
Kineman R, Frawley L. Secretory characteristics and phenotypic plasticity of growth hormone and prolactin-producing cell lines. J Endocrinol 1994; 140: 455–463.
Chatellard-Causse C, Blot B, Cristina N, Torch S, Missotten M, Sadoul R. Alix (ALG-2-interacting protein X) a protein involved in apoptosis binds to endophilins and induces cytoplasmic vacuolization. J Biol Chem 2002; 277: 29108–29115.
Torriglia A, Perani P, Brossas J, et al. L-DNaseII, a molecule that links proteases and endonucleases in apoptosis, derives from the ubiquitous serpin Leukocyte Elastase Inhibitor. Mol Cell Biol. 1998; 18: 3612–3619.
Altairac S, Chaudun I, Courtois I, Torriglia A. Elastase is not required for L-DNaseII activation during apoptosis in developing chicken neural retina. Neurosci Lett 2001; 303: 41–44.
Altairac S, Wright S, Courtois Y, Torriglia A. L-DNaseII activation by the 24 kDa apoptotic protease (AP24) in TNF-alpha-induced apoptosis. Cell Death Differ 2003; 10: 1109–1111.
Perani P, Zeggari S, Torriglia A, Courtois I. Mutations on the hinge region of leukocyte elastase inhibitor determine the loss of inhibitory function. Biochem Biohys Res Comm 2000; 274: 841–844.
Belmokhtar C, Torriglia A, Counis M, Courtois Y, Jacquemin-Sablon A, Ségal-Bendirdjian E. Nuclear translocation of a Leukocyte Elastase Inhibitor/Elastase complex during staurosporine-induced apoptosis: Role in the generation of nuclear L-DNaseII activity. Exp Cell Res 2000; 254: 99– 109.
Lockshin R, Zakeri Z. Programmed cell death and apoptosis: Origins of the theory. Nat Rev Mol Cell Biol 2001; 2: 545–550.
Lockshin R, Zakeri Z. Caspase-independent cell deaths. Curr Opinion Cell Biol 2002; 14: 727–733.
Lockshin R, Zakeri Z. Apoptosis, autophagy, and more. Int J Biochem & Cell Biol. 2004; 36: 2405–2419.
Lockshin R, Osborne B, Zakeri Z. Cell death in the third millennium. Cell Death Differ. 2000; 7: 2–7.
Chahory S, Padron L, Courtois I, Torriglia A. The LEI/L-DNase II pathway is activated in light-induced retinal degeneration in rats. Neurosci Lett. 2004; 367: 205–209.
Aanestad M, Rotnes J, Torjesen P, Haug E, Sand O, Bjoro T. Epidermal growth factor stimulates the prolactin synthesis and secretion in rat pituitary cells in culture (GH4C1 cells) by increasing the intracellular concentration of free calcium. Acta Endocrinol (Copenh) 1993; 128: 361–366.
Tsukada T, Watanabe M, Yamashima T. Implications of CAD and DNaseII in ischemic neuronal necrosis for the primate hippocampus. J Neurochem 2001; 79: 1196–1206.
Missotten M, Nichols A, Rieger K, Sadoul R. Alix, a novel mouse protein undergoing calcium-dependent interaction with the apoptosis-linked gene-2 (ALG-2) protein. Cell Death Differ. 1999; 6: 124–129.
Vito P, Pellegrini L, Guiet C, D'Adamio L. Cloning of AIP1, a novel protein that associates with the apoptosis-linked gene ALG-2 in Ca2+-dependent reaction. J Biol Chem 1999; 274: 1533–1540.
Schmidt M, Hoeller D, Yu J, et al. Alix/AIP1 antagonizes epidermal growth factor receptor downregulation by the Cbl-SETA/CIN85 complex. Mol Cell Biol 2004; 24: 8981–8993.
Birman P, Michard M, Li J, Peillon F, Bression D. Epidermal growth factor-binding sites, present in normal human and rat pituitaries, are absent in human pituitary adenomas. J Clin Endocrinol Metab 1987; 65: 275–281.
Author information
Authors and Affiliations
Corresponding author
Additional information
This work was supported by fellowships from Fondation pour la Recherche medicale (FRM: FDT20030627283), from Association pour la Recherche contre le Cancer (ARC: ML/MLD/CM-A04/4) to JF and Retina France to AT and LP.
These two authors equally contributed to this work.
Rights and permissions
About this article
Cite this article
Fombonne, J., Padrón, L., Enjalbert, A. et al. A novel paraptosis pathway involving LEI/L-DNaseII for EGF-induced cell death in somato-lactotrope pituitary cells. Apoptosis 11, 367–375 (2006). https://doi.org/10.1007/s10495-006-4568-6
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10495-006-4568-6