Identification of c-Jun as bcl-2 Transcription Factor in Human Uterine Endometrium

We describe the application of the biomolecular interaction (BIA) technique to detection of the interaction between protein (e.g., c-Jun) and DNA (e.g., two AP-1 motifs from bcl-2 promoter), compared with immunohistochemistry (IHC) of c-Jun. The specific binding assay for the interaction of c-Jun and activating protein-1 (AP-1) motifs was performed using a Biacore 2000 system. Intense immunoreactivity of c-Jun in glandular cells of the human uterine endometrium was observed in the proliferative phase, while c-Jun in stromal cells was expressed throughout the menstrual cycle. In contrast to the IHC of c-Jun, the specific binding of c-Jun to two separate AP-1 motifs in the bcl-2 promoter region was detected only in nuclear extracts of glandular cells, but not in stromal cells, during the proliferative phase. These results indicate that, while transmitting various signals, c-Jun enhances the transcription level of bcl-2, which in turn keeps glandular cells alive and proliferating in normal human endometrium during the proliferative phase. Moreover, the method involving real-time biomolecular interactions such as DNA–protein binding is novel for the study of transcription factors when combined with IHC.

[1]  M. Negroni,et al.  Real time measurements of elongation by a reverse transcriptase using surface plasmon resonance. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[2]  T. Kerppola,et al.  Close encounters of many kinds: Fos-Jun interactions that mediate transcription regulatory specificity , 2001, Oncogene.

[3]  N. Fusenig,et al.  c-Jun and JunB Antagonistically Control Cytokine-Regulated Mesenchymal–Epidermal Interaction in Skin , 2000, Cell.

[4]  D. Latchman,et al.  Bcl-2 Transcription from the Proximal P2 Promoter Is Activated in Neuronal Cells by the Brn-3a POU Family Transcription Factor* , 1998, The Journal of Biological Chemistry.

[5]  J. Guillet,et al.  Analysis of interactions between huGATA-3 transcription factor and three GATA regulatory elements of HIV-1 long terminal repeat, by surface plasmon resonance. , 1997, Analytical biochemistry.

[6]  E. Rutanen,et al.  Cellular Localization of c-Jun Messenger Ribonucleic Acid and Protein and Their Relation to the Proliferation Marker Ki-67 in the Human Endometrium , 1998 .

[7]  Y. Akao,et al.  Cyclic bcl-2 gene expression in human uterine endometrium during menstrual cycle , 1994, The Lancet.

[8]  Hans van Dam,et al.  Distinct roles of Jun : Fos and Jun : ATF dimers in oncogenesis , 2001, Oncogene.

[9]  J. Erickson,et al.  Real‐time DNA binding measurements of the ETSl recombinant oncoproteins reveal significant kinetic differences between the p42 and p51 isoforms , 1994, Protein science : a publication of the Protein Society.

[10]  M. Olive,et al.  CRE DNA binding proteins bind to the AP-1 target sequence and suppress AP-1 transcriptional activity in mouse keratinocytes , 1999, Oncogene.

[11]  Z. Oltvai,et al.  Bcl-2 gene family and the regulation of programmed cell death. , 1994, Cold Spring Harbor symposia on quantitative biology.

[12]  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.

[13]  John Calvin Reed,et al.  Mechanisms of Transcriptional Activation of bcl-2Gene Expression by 17β-Estradiol in Breast Cancer Cells* , 1999, The Journal of Biological Chemistry.

[14]  P. Vogt,et al.  Avian sarcoma virus 17 carries the jun oncogene. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[15]  G. Palumbo,et al.  17β-Estradiol Inhibits Apoptosis in MCF-7 Cells, Inducing bcl-2 Expression via Two Estrogen-Responsive Elements Present in the Coding Sequence , 2000, Molecular and Cellular Biology.

[16]  C. Vaquero,et al.  Real-time study of interactions between a composite DNA regulatory region (HIV-1 LTR NRE) and several transcription factors of nuclear extracts. , 1999, Biochemical and biophysical research communications.

[17]  R. Fisher,et al.  BIAcore for macromolecular interaction. , 1998, Current opinion in biotechnology.

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

[19]  Y. Tsujimoto,et al.  Involvement of the bcl-2 gene in human follicular lymphoma. , 1985, Science.

[20]  Yuko Ito,et al.  Fas-mediated apoptosis in human lens epithelial cells of cataracts associated with diabetic retinopathy , 2002, Medical Electron Microscopy.

[21]  Yuko Ito,et al.  The activation of caspase-3 and DNA fragmentation in B cells phagocytosed by macrophages , 2003, Medical Electron Microscopy.

[22]  Frank Hilberg,et al.  Functions of c-Jun in Liver and Heart Development , 1999, The Journal of cell biology.

[23]  M. Karin,et al.  AP-1 in cell proliferation and survival , 2001, Oncogene.

[24]  M. Karin,et al.  The role of Jun, Fos and the AP-1 complex in cell-proliferation and transformation. , 1991, Biochimica et biophysica acta.