Identification of DIABLO, a Mammalian Protein that Promotes Apoptosis by Binding to and Antagonizing IAP Proteins

To identify proteins that bind mammalian IAP homolog A (MIHA, also known as XIAP), we used coimmuno-precipitation and 2D immobilized pH gradient/SDS PAGE, followed by electrospray ionization tandem mass spectrometry. DIABLO (direct IAP binding protein with low pI) is a novel protein that can bind MIHA and can also interact with MIHB and MIHC and the baculoviral IAP, OpIAP. The N-terminally processed, IAP-interacting form of DIABLO is concentrated in membrane fractions in healthy cells but released into the MIHA-containing cytosolic fractions upon UV irradiation. As transfection of cells with DIABLO was able to counter the protection afforded by MIHA against UV irradiation, DIABLO may promote apoptosis by binding to IAPs and preventing them from inhibiting caspases.

[1]  Xiaodong Wang,et al.  Smac, a Mitochondrial Protein that Promotes Cytochrome c–Dependent Caspase Activation by Eliminating IAP Inhibition , 2000, Cell.

[2]  H. Steller,et al.  The head involution defective gene of Drosophila melanogaster functions in programmed cell death. , 1995, Genes & development.

[3]  Sharad Kumar,et al.  DRONC, an ecdysone-inducible Drosophila caspase. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[4]  W. Kaiser,et al.  The Drosophila inhibitor of apoptosis D‐IAP1 suppresses cell death induced by the caspase drICE , 1998, FEBS letters.

[5]  V. Dixit,et al.  Caspase-9, Bcl-XL, and Apaf-1 Form a Ternary Complex* , 1998, The Journal of Biological Chemistry.

[6]  R. Moritz,et al.  Application of capillary reversed-phase high-performance liquid chromatography to high-sensitivity protein sequence analysis. , 1992, Journal of chromatography.

[7]  Yuanming Hu,et al.  Bcl-XL interacts with Apaf-1 and inhibits Apaf-1-dependent caspase-9 activation. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[8]  B. Hay,et al.  A cloning method to identify caspases and their regulators in yeast: identification of Drosophila IAP1 as an inhibitor of the Drosophila caspase DCP-1. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[9]  R. Rasmussen,et al.  Capillary column chromatography improves sample preparation for mass spectrometric analysis: Complete characterization of human α‐enolase from two‐dimensional gels following in situ proteolytic digestion , 1998, Electrophoresis.

[10]  Mike Rothe,et al.  The TNFR2-TRAF signaling complex contains two novel proteins related to baculoviral inhibitor of apoptosis proteins , 1995, Cell.

[11]  Margot Thome,et al.  Inhibition of death receptor signals by cellular FLIP , 1997, Nature.

[12]  R. Moritz,et al.  S‐Pyridylethylation of intact polyacrylamide gels and in situ digestion of electrophoretically separated proteins: A rapid mass spectrometric method for identifying cysteine‐containing peptides , 1996, Electrophoresis.

[13]  C. Thompson,et al.  A conserved family of cellular genes related to the baculovirus iap gene and encoding apoptosis inhibitors. , 1996, The EMBO journal.

[14]  D. Vaux,et al.  Cloning and expression of apoptosis inhibitory protein homologs that function to inhibit apoptosis and/or bind tumor necrosis factor receptor-associated factors. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[15]  K. Tamai,et al.  Suppression of apoptosis in mammalian cells by NAIP and a related family of IAP genes , 1996, Nature.

[16]  R. J. Clem,et al.  An apoptosis-inhibiting baculovirus gene with a zinc finger-like motif , 1993, Journal of virology.

[17]  S. Korsmeyer,et al.  Cell Death in Development , 1999, Cell.

[18]  J C Reed,et al.  IAPs block apoptotic events induced by caspase‐8 and cytochrome c by direct inhibition of distinct caspases , 1998, The EMBO journal.

[19]  S. Cory,et al.  Bcl-2 family members do not inhibit apoptosis by binding the caspase activator Apaf-1. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[20]  S. Seshagiri,et al.  Baculovirus inhibitors of apoptosis (IAPs) block activation of Sf-caspase-1. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[21]  H. Müller,et al.  The Drosophila Caspase Inhibitor DIAP1 Is Essential for Cell Survival and Is Negatively Regulated by HID , 1999, Cell.

[22]  P. Roepstorff,et al.  Proposal for a common nomenclature for sequence ions in mass spectra of peptides. , 1984, Biomedical mass spectrometry.

[23]  Gerald M. Rubin,et al.  Drosophila homologs of baculovirus inhibitor of apoptosis proteins function to block cell death , 1995, Cell.

[24]  D. Vaux,et al.  Inhibition of interleukin 1 beta-converting enzyme-mediated apoptosis of mammalian cells by baculovirus IAP. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[25]  H. Steller,et al.  Genetic control of programmed cell death in Drosophila. , 1994, Science.

[26]  David Wallach,et al.  Involvement of MACH, a Novel MORT1/FADD-Interacting Protease, in Fas/APO-1- and TNF Receptor–Induced Cell Death , 1996, Cell.

[27]  Xiaodong Wang,et al.  Apaf-1, a Human Protein Homologous to C. elegans CED-4, Participates in Cytochrome c–Dependent Activation of Caspase-3 , 1997, Cell.

[28]  R. Moritz,et al.  A two‐dimensional gel database of human colon carcinoma proteins , 1997, Electrophoresis.

[29]  Guy S. Salvesen,et al.  X-linked IAP is a direct inhibitor of cell-death proteases , 1997, Nature.

[30]  Junying Yuan,et al.  Human ICE/CED-3 Protease Nomenclature , 1996, Cell.

[31]  J. Abrams,et al.  grim, a novel cell death gene in Drosophila. , 1996, Genes & development.

[32]  D. Vaux,et al.  Inhibition of apoptosis and clonogenic survival of cells expressing crmA variants: optimal caspase substrates are not necessarily optimal inhibitors , 1999, The EMBO journal.

[33]  D. Scherman,et al.  A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[34]  Matthias Mann,et al.  FLICE, A Novel FADD-Homologous ICE/CED-3–like Protease, Is Recruited to the CD95 (Fas/APO-1) Death-Inducing Signaling Complex , 1996, Cell.