Induction and endoplasmic reticulum location of BIK/NBK in response to apoptotic signaling by E1A and p53

A DNA microarray analysis identified the BH3-only BCL-2 family member, BIK/NBK, as a transcript that is upregulated during induction of apoptosis by oncogenic E1A. E1A depended on wild-type p53 to induce BIK and activate the death program. Further, p53 independently induced BIK RNA and protein, and BIK alone stimulated cell death in p53-null cells, dependent on the activation of caspases. BIK function, however, was abrogated by a disabling point mutation within the BH3 domain. Collectively, these results argue that BIK is a downstream apoptotic effector of p53 in response to a physiological p53-mediated death stimulus provided by E1A. Elevated BCL-2 functioned downstream of p53 and BIK induction to inhibit the E1A death pathway, with the ratio of anti-apoptotic BCL-2 and pro-apoptotic BIK determining cell death or survival in E1A-expressing cells. Cells expressing BCL-2 or treated with the pan caspase inhibitor, zVAD-fmk, allowed accumulation of high levels of cytotoxic BIK compared to control cells. Of note, a significant fraction of either ectopic or endogenous BIK was found associated with the endoplasmic reticulum, suggesting that this organelle, in addition to mitochondria, may be a target of BIK function.

[1]  S. Lowe,et al.  Stabilization of the p53 tumor suppressor is induced by adenovirus 5 E1A and accompanies apoptosis. , 1993, Genes & development.

[2]  J. Howe,et al.  Induction of the cell cycle in baby rat kidney cells by adenovirus type 5 E1A in the absence of E1B and a possible influence of p53 , 1993, Journal of virology.

[3]  G. Chinnadurai,et al.  Functional Dissection of the Pro-apoptotic Protein Bik , 1997, The Journal of Biological Chemistry.

[4]  K. Vousden,et al.  PUMA, a novel proapoptotic gene, is induced by p53. , 2001, Molecular cell.

[5]  S. Lowe.  Activation of p53 by oncogenes. , 1999, Endocrine-related cancer.

[6]  W. Zong,et al.  BH3-only proteins that bind pro-survival Bcl-2 family members fail to induce apoptosis in the absence of Bax and Bak. , 2001, Genes & development.

[7]  P. Krammer,et al.  CD95's deadly mission in the immune system , 2000, Nature.

[8]  V. Dixit,et al.  Bik and Bak Induce Apoptosis Downstream of CrmA but Upstream of Inhibitor of Apoptosis* , 1997, The Journal of Biological Chemistry.

[9]  Junying Yuan,et al.  Caspase-12 mediates endoplasmic-reticulum-specific apoptosis and cytotoxicity by amyloid-β , 2000, Nature.

[10]  J. Growdon,et al.  Presenilin 1 Protein Directly Interacts with Bcl-2* , 1999, The Journal of Biological Chemistry.

[11]  E. Alnemri,et al.  E1A-induced Processing of Procaspase-8 Can Occur Independently of FADD and Is Inhibited by Bcl-2* , 1998, The Journal of Biological Chemistry.

[12]  P. Branton,et al.  p28 Bap31, a Bcl-2/Bcl-XL- and Procaspase-8–associated Protein in the Endoplasmic Reticulum , 1997, The Journal of cell biology.

[13]  M. Lutter,et al.  Biochemical pathways of caspase activation during apoptosis. , 1999, Annual review of cell and developmental biology.

[14]  M. Hengartner.  The biochemistry of apoptosis , 2000, Nature.

[15]  L. Pellegrini,et al.  Interaction of Alzheimer's presenilin-1 and presenilin-2 with Bcl-X(L). A potential role in modulating the threshold of cell death. , 1999, The Journal of biological chemistry.

[16]  S. Srinivasula,et al.  Blk, a BH3-containing Mouse Protein That Interacts with Bcl-2 and Bcl-xL, Is a Potent Death Agonist* , 1998, The Journal of Biological Chemistry.

[17]  Y. Akao,et al.  Multiple subcellular localization of bcl-2: detection in nuclear outer membrane, endoplasmic reticulum membrane, and mitochondrial membranes. , 1994, Cancer research.

[18]  John Calvin Reed,et al.  Tumor suppressor p53 is a direct transcriptional activator of the human bax gene , 1995, Cell.

[19]  M. Vu,et al.  Proteasomes Modulate Balance Among Proapoptotic and Antiapoptotic Bcl-2 Family Members and Compromise Functioning of the Electron Transport Chain in Leukemic Cells1 , 2001, The Journal of Immunology.

[20]  I. Krantz,et al.  KILLER/DR5 is a DNA damage–inducible p53–regulated death receptor gene , 1997, Nature Genetics.

[21]  C. Prives,et al.  The p53 pathway , 1999, The Journal of pathology.

[22]  Yunping Lin,et al.  Pidd, a new death-domain–containing protein, is induced by p53 and promotes apoptosis , 2000, Nature Genetics.

[23]  S. Korsmeyer,et al.  The adenovirus E1A proteins induce apoptosis, which is inhibited by the E1B 19-kDa and Bcl-2 proteins. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[24]  S. Korsmeyer,et al.  Regulated Targeting of BAX to Mitochondria , 1998, The Journal of cell biology.

[25]  G. Evan,et al.  Caspase 8: The killer you can't live without , 2000, Nature Medicine.

[26]  M. Ljungman.  Dial 9-1-1 for p53: mechanisms of p53 activation by cellular stress. , 2000, Neoplasia.

[27]  S. Chiou,et al.  Interleukin 1 beta converting enzyme-like proteases are essential for p53-mediated transcriptionally dependent apoptosis. , 1997, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[28]  S. Korsmeyer,et al.  Proapoptotic BAX and BAK: A Requisite Gateway to Mitochondrial Dysfunction and Death , 2001, Science.

[29]  F. Graham,et al.  A simple technique for the rescue of early region I mutations into infectious human adenovirus type 5. , 1988, Virology.

[30]  M. Takeda,et al.  A novel protein, RTN-xS, interacts with both Bcl-xL and Bcl-2 on endoplasmic reticulum and reduces their anti-apoptotic activity , 2000, Oncogene.

[31]  S. Verma,et al.  Phosphorylation of the Pro-apoptotic Protein BIK , 2001, The Journal of Biological Chemistry.

[32]  K. Kinzler,et al.  PUMA induces the rapid apoptosis of colorectal cancer cells. , 2001, Molecular cell.

[33]  F. Graham,et al.  An efficient and flexible system for construction of adenovirus vectors with insertions or deletions in early regions 1 and 3. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[34]  E. White,et al.  Induction of apoptosis by human Nbk/Bik, a BH3-containing protein that interacts with E1B 19K , 1996, Molecular and cellular biology.

[35]  P. Branton,et al.  Accumulation of p53 induced by the adenovirus E1A protein requires regions involved in the stimulation of DNA synthesis , 1997, Journal of virology.

[36]  G. Shore,et al.  Regulation of apoptosis by E1A and Myc oncoproteins. , 2000, Critical reviews in eukaryotic gene expression.

[37]  E. White,et al.  Wild-type p53 mediates apoptosis by E1A, which is inhibited by E1B. , 1993, Genes & development.

[38]  E. White,et al.  Life, death, and the pursuit of apoptosis. , 1996, Genes & development.

[39]  G. G. Stokes.  "J." , 1890, The New Yale Book of Quotations.

[40]  Yusuke Nakamura,et al.  p53AIP1, a Potential Mediator of p53-Dependent Apoptosis, and Its Regulation by Ser-46-Phosphorylated p53 , 2000, Cell.

[41]  John Calvin Reed,et al.  Investigation of the subcellular distribution of the bcl-2 oncoprotein: residence in the nuclear envelope, endoplasmic reticulum, and outer mitochondrial membranes. , 1993, Cancer research.

[42]  G. Shore,et al.  Caspase-Resistant BAP31 Inhibits Fas-Mediated Apoptotic Membrane Fragmentation and Release of Cytochrome cfrom Mitochondria , 2000, Molecular and Cellular Biology.

[43]  S. Korsmeyer,et al.  The combined functions of proapoptotic Bcl-2 family members bak and bax are essential for normal development of multiple tissues. , 2000, Molecular cell.

[44]  K. Vousden,et al.  Minireviewp 53 : Death Star able to induce the defensive p 53 response to oncogene , 2000 .

[45]  A. Strasser,et al.  BH3-Only Proteins—Essential Initiators of Apoptotic Cell Death , 2000, Cell.

[46]  S. Korsmeyer,et al.  BCL-2 family members and the mitochondria in apoptosis. , 1999, Genes & development.