Notch2 is involved in the overexpression of CD23 in B-cell chronic lymphocytic leukemia.
Members of the Notch family encode transmembrane receptors that modulate differentiation, proliferation, and apoptotic programs of many precursor cells, including hematopoietic progenitors. Stimulation of Notch causes cleavage followed by translocation of the intracellular domain (NotchIC) to the nucleus, where it activates transcription of CBF1 responsive genes. The aim of this study was to elucidate the mechanisms leading to the overexpression of CD23, a striking feature of B-cell chronic lymphocytic leukemia (B-CLL) cells. By electrophoretic mobility shift assays, we identified a transcription factor complex (C1) that binds sequence specific to one known and 4 newly identified putative CBF1 recognition sites in the CD23a core promoter region. With the use of Epstein-Barr virus (EBV)-infected B cells as a model for CBF1 mediated CD23a expression, C1 was found to be EBV inducible. Supershift assays revealed that the nuclear form of Notch2 is a component of C1 in B-CLL cells, supporting a model in which NotchIC activates transcription by binding to CBF1 tethered to DNA. Transient transfection of REH pre-B cells with an activated form of Notch2 induced endogenous CD23a, confirming that CD23a is a target gene of Notch2 signaling. Finally, reverse transcription-polymerase chain reaction and kinetic analysis demonstrated that the Notch2 oncogene is not only overexpressed in B-CLL cells but might also be related to the failure of apoptosis characteristic for this disease. In conclusion, these data suggest that deregulation of Notch2 signaling is involved in the aberrant expression of CD23 in B-CLL.
Hubmann
Rainer
R
Department of Hematology, Clinic of Internal Medicine I, University of Vienna, Vienna, Austria.
Schwarzmeier
Josef D
JD
Shehata
Medhat
M
Hilgarth
Martin
M
Duechler
Markus
M
Dettke
Markus
M
Berger
Rudolf
R
D017209:Apoptosis; D001402:B-Lymphocytes / Q000378:metabolism; D001483:Base Sequence; D002460:Cell Line; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012334:RNA, Neoplasm / Q000096:biosynthesis; D051883:Receptor, Notch2; D011956:Receptors, Cell Surface / Q000235:genetics / Q000502:physiology*; D017455:Receptors, IgE / Q000235:genetics* / Q000378:metabolism; D020218:Response Elements; D015533:Transcriptional Activation; D015854:Up-Regulation
D004268:DNA-Binding Proteins; C497764:NOTCH2 protein, human; D012334:RNA, Neoplasm; D051883:Receptor, Notch2; D011956:Receptors, Cell Surface; D017455:Receptors, IgE