CTCF functions as a critical regulator of cell-cycle arrest and death after ligation of the B cell receptor on immature B cells

The WEHI 231 B cell lymphoma is used as a model of self-tolerance by clonal deletion because B cell receptor (BCR) ligation results in apoptosis. Two critical events precede cell death: an early rise and fall in expression of MYC and cell-cycle arrest associated with enhanced expression of p21, p27, and p53. CTCF is a transcription factor identified as a repressor of MYC recently shown to cause cell growth inhibition. The present studies demonstrate that BCR ligation of WEHI 231 as well as of normal immature B cells greatly increased expression of CTCF in association with down-regulation of MYC followed by growth arrest and cell death. Conditional expression of CTCF in WEHI 231 mimicked BCR ligation with activated cells showing repressed expression of MYC, enhanced expression of p27, p21, p53, and p19ARF, and inhibition of cell growth and induction of apoptosis. In keeping with a central role for CTCF in control of B cell death, conditional expression of a CTCF antisense construct in WEHI 231 resulted in inhibition of p27, p21, p53, and p19ARF in association with enhanced expression of MYC. Activation of the endogenous CTCF locus by BCR ligation was also mimicked by three other routes to apoptotic death in WEHI 231: inhibition of the phosphoinositide 3-kinase or mTOR/FRAP signaling cascades and treatment with transforming growth factor (TGF)-β. Rapid activation of CTCF by BCR ligation or treatment with TGF-β was suppressed by ligation of CD40. These results demonstrate that CTCF is a common determinant to different pathways of death signaling in immature B cells.

[1]  David Baltimore,et al.  Cooperation of multiple signaling pathways in CD40-regulated gene expression in B lymphocytes , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[2]  R Ohlsson,et al.  CTCF is a uniquely versatile transcription regulator linked to epigenetics and disease. , 2001, Trends in genetics : TIG.

[3]  P. Neiman,et al.  Cell growth inhibition by the multifunctional multivalent zinc-finger factor CTCF. , 2001, Cancer research.

[4]  D. W. Kim,et al.  Repression of transcription of the p27Kip1 cyclin-dependent kinase inhibitor gene by c-Myc , 2001, Oncogene.

[5]  P. Neiman,et al.  Functional Phosphorylation Sites in the C-Terminal Region of the Multivalent Multifunctional Transcriptional Factor CTCF , 2001, Molecular and Cellular Biology.

[6]  R. Parsons,et al.  PTEN: life as a tumor suppressor. , 2001, Experimental cell research.

[7]  D. Scott,et al.  Role of Phosphatidylinositol 3-Kinase in Anti-IgM- and Anti-IgD-Induced Apoptosis in B Cell Lymphomas1 2 , 2001, The Journal of Immunology.

[8]  J. Massagué,et al.  Defective repression of c-myc in breast cancer cells: A loss at the core of the transforming growth factor beta growth arrest program. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[9]  Chen-feng Qi,et al.  Genomic organisation and expression of BCL6 in murine B-cell lymphomas. , 2000, Leukemia research.

[10]  Victor V Lobanenkov,et al.  Functional association of CTCF with the insulator upstream of the H19 gene is parent of origin-specific and methylation-sensitive , 2000, Current Biology.

[11]  Shirley M. Tilghman,et al.  CTCF mediates methylation-sensitive enhancer-blocking activity at the H19/Igf2 locus , 2000, Nature.

[12]  G. Felsenfeld,et al.  Methylation of a CTCF-dependent boundary controls imprinted expression of the Igf2 gene , 2000, Nature.

[13]  L. Lau,et al.  IPTG-inducible episomal expression system for exogenous genes in primate cells. , 2000, BioTechniques.

[14]  L. Chin,et al.  Differential effects of the widely expressed dMax splice variant of Max on E-box vs initiator element-mediated regulation by c-Myc , 1999, Oncogene.

[15]  Kou-Juey Wu,et al.  Coordinated regulation of iron-controlling genes, H-ferritin and IRP2, by c-MYC. , 1999, Science.

[16]  V. Kouskoff,et al.  Antigens Varying in Affinity for the B Cell Receptor Induce Differential B Lymphocyte Responses , 1998, The Journal of experimental medicine.

[17]  C. Sherr,et al.  Tumor surveillance via the ARF-p53 pathway. , 1998, Genes & development.

[18]  Kevin Ryan,et al.  The alternative product from the human CDKN2A locus, p14ARF, participates in a regulatory feedback loop with p53 and MDM2 , 1998, The EMBO journal.

[19]  M. West,et al.  Translational induction of the c-myc oncogene via activation of the FRAP/TOR signalling pathway , 1998, Oncogene.

[20]  F. Zindy,et al.  Functional and physical interactions of the ARF tumor suppressor with p53 and Mdm2. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[21]  M. Wu,et al.  Roles of the Tumor Suppressor p53 and the Cyclin-dependent Kinase Inhibitor p21WAF1/CIP1 in Receptor-mediated Apoptosis of WEHI 231 B Lymphoma Cells , 1998, The Journal of experimental medicine.

[22]  Ken Chen,et al.  The Ink4a Tumor Suppressor Gene Product, p19Arf, Interacts with MDM2 and Neutralizes MDM2's Inhibition of p53 , 1998, Cell.

[23]  A. Levine.  p53, the Cellular Gatekeeper for Growth and Division , 1997, Cell.

[24]  M. Arsura,et al.  TGFβ1 Inhibits NF-κB/Rel Activity Inducing Apoptosis of B Cells: Transcriptional Activation of IκBα , 1996 .

[25]  P. Neiman,et al.  An exceptionally conserved transcriptional repressor, CTCF, employs different combinations of zinc fingers to bind diverged promoter sequences of avian and mammalian c-myc oncogenes , 1996, Molecular and cellular biology.

[26]  D. Scott,et al.  Role of cyclin A and p27 in anti-IgM induced G1 growth arrest of murine B-cell lymphomas. , 1996, Molecular biology of the cell.

[27]  M. Arsura,et al.  Role of Rel-related factors in control of c-myc gene transcription in receptor-mediated apoptosis of the murine B cell WEHI 231 line , 1995, The Journal of experimental medicine.

[28]  D. Green,et al.  Lymphoma models for B cell activation and tolerance. X. Anti-mu- mediated growth arrest and apoptosis of murine B cell lymphomas is prevented by the stabilization of myc , 1994, The Journal of experimental medicine.

[29]  J. Trent,et al.  WAF1, a potential mediator of p53 tumor suppression , 1993, Cell.

[30]  S. Camper,et al.  Receptor editing: an approach by autoreactive B cells to escape tolerance , 1993, The Journal of experimental medicine.

[31]  D. Nemazee,et al.  Receptor editing in self-reactive bone marrow B cells , 1993, The Journal of experimental medicine.

[32]  Victor V Lobanenkov,et al.  A novel sequence-specific DNA binding protein which interacts with three regularly spaced direct repeats of the CCCTC-motif in the 5'-flanking sequence of the chicken c-myc gene. , 1990, Oncogene.

[33]  G. Klaus,et al.  Anti‐immunoglobulin antibodies induce apoptosis in immature B cell lymphomas , 1990, European journal of immunology.

[34]  P. Cazenave,et al.  Anti‐immunoglobulins induce death by apoptosis in WEHI‐231 B lymphoma cells , 1990, European journal of immunology.

[35]  S. Smith‐Gill,et al.  Altered immunoglobulin expression and functional silencing of self-reactive B lymphocytes in transgenic mice , 1988, Nature.

[36]  D. Scott,et al.  Lymphoma models for B cell activation and tolerance IV. Growth inhibition by anti‐Ig of CH31 and CH33 B lymphoma cells , 1986, European journal of immunology.

[37]  G. Sonenshein,et al.  Specific regulation of c-myc oncogene expression in a murine B-cell lymphoma. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[38]  A. Boyd,et al.  Clonal anergy: the universally anergic B lymphocyte. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[39]  A. Boyd,et al.  The regulation of growth and differentiation of a murine B cell lymphoma. II. The inhibition of WEHI 231 by anti-immunoglobulin antibodies. , 1981, Journal of immunology.