The Pro-Oncoprotein EWS (Ewing’s Sarcoma Protein) Interacts with the Brn-3a POU Transcription Factor and Inhibits its Ability Activate Transcription

The Brn-3a POU family transcription factor is able to induce the expression of a number of neuronally-expressed genes as well as to enhance neuronal differentiation and inhibit apoptosis. Many of these effects are mediated by the C-terminal POU domain of Brn-3a which acts both as a DNA binding domain and a transcriptional activation domain. To identify the mechanisms by which this domain acts, we carried out a yeast two hybrid assay to identify proteins which interact with it. We show that both full length Brn-3a and the isolated POU domain interact with the EWS transcription factor and its oncogenic derivative EWS-Fli1. Moreover, EWS can block Brn-3a-mediated activation of the Bcl-x promoter whereas this effect is lost in EWS-Fli1. The significance of this novel interaction is discussed in terms of the manner in which Brn-3a regulates its target promoters and the mechanism of oncogenic transformation by EWS-Fli1.

[1]  D. Latchman,et al.  Brn-3a Activates the Expression of Bcl-xL and Promotes Neuronal Survival in Vivo as Well as in Vitro , 2001, Molecular and Cellular Neuroscience.

[2]  D. Latchman,et al.  The BRN-3A Transcription Factor Protects Sensory but Not Sympathetic Neurons from Programmed Cell Death/Apoptosis* , 2001, The Journal of Biological Chemistry.

[3]  K K Li,et al.  Transcriptional Activation by the Ewing's Sarcoma (EWS) Oncogene Can Be Cis-repressed by the EWS RNA-binding Domain* , 2000, The Journal of Biological Chemistry.

[4]  P. Sorensen,et al.  EWS/ETS fusion genes induce epithelial and neuroectodermal differentiation in NIH 3T3 fibroblasts. , 1999, Laboratory investigation; a journal of technical methods and pathology.

[5]  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.

[6]  H. Kovar,et al.  Ewing's sarcoma and peripheral primitive neuroectodermal tumors after their genetic union. , 1998, Current opinion in oncology.

[7]  Olivier Delattre,et al.  EWS, but Not EWS-FLI-1, Is Associated with Both TFIID and RNA Polymerase II: Interactions between Two Members of the TET Family, EWS and hTAFII68, and Subunits of TFIID and RNA Polymerase II Complexes , 1998, Molecular and Cellular Biology.

[8]  P. Sorensen,et al.  Absence of detectable EWS/FLI1 expression after therapy-induced neural differentiation in Ewing sarcoma. , 1998, Human pathology.

[9]  M. Schwartz,et al.  Coordinate Induction of the Three Neurofilament Genes by the Brn-3a Transcription Factor* , 1997, The Journal of Biological Chemistry.

[10]  D. Latchman,et al.  A single residue within the homeodomain of the Brn‐3 POU family transcription factors determines whether they activate or repress the SNAP‐25 promoter , 1997, Neuroreport.

[11]  M. Rosenfeld,et al.  POU domain family values: flexibility, partnerships, and developmental codes. , 1997, Genes & development.

[12]  M. Ouchida,et al.  Inhibition of apoptosis by normal and aberrant Fli-1 and erg proteins involved in human solid tumors and leukemias , 1997, Oncogene.

[13]  K. Tanaka,et al.  EWS-Fli1 antisense oligodeoxynucleotide inhibits proliferation of human Ewing's sarcoma and primitive neuroectodermal tumor cells. , 1997, The Journal of clinical investigation.

[14]  D. Latchman,et al.  The Brn-3a transcription factor induces neuronal process outgrowth and the coordinate expression of genes encoding synaptic proteins , 1997, Molecular and cellular biology.

[15]  D. Latchman,et al.  Differential regulation of genes encoding synaptic proteins by members of the Brn-3 subfamily of POU transcription factors. , 1996, Brain research. Molecular brain research.

[16]  P. Sawchenko,et al.  Requirement for Brn-3.0 in differentiation and survival of sensory and motor neurons , 1996, Nature.

[17]  J. Nathans,et al.  Targeted deletion of the mouse POU domain gene Brn-3a causes selective loss of neurons in the brainstem and trigeminal ganglion, uncoordinated limb movement, and impaired suckling. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[18]  D. Latchman,et al.  The functionally antagonistic POU family transcription factors Brr‐3a and Brn‐3b show opposite changes in expression during the growth arrest and differentiation of human neuroblastoma cells , 1996, International journal of cancer.

[19]  D. Latchman,et al.  A Single Amino Acid Change Converts an Inhibitory Transcription Factor into an Activator (*) , 1996, The Journal of Biological Chemistry.

[20]  E. Turner,et al.  Brn-3.0 expression identifies early post-mitotic CNS neurons and sensory neural precursors , 1995, Mechanisms of Development.

[21]  M. Ouchida,et al.  Loss of tumorigenicity of Ewing's sarcoma cells expressing antisense RNA to EWS-fusion transcripts. , 1995, Oncogene.

[22]  T. Möröy,et al.  Regulation of Neurite Outgrowth and SNAP-25 Gene Expression by the Brn-3a Transcription Factor (*) , 1995, The Journal of Biological Chemistry.

[23]  T. Möröy,et al.  Activation of the α-Internexin Promoter by the Brn-3a Transcription Factor Is Dependent on the N-terminal Region of the Protein (*) , 1995, The Journal of Biological Chemistry.

[24]  M. Ouchida,et al.  The EWS gene, involved in Ewing family of tumors, malignant melanoma of soft parts and desmoplastic small round cell tumors, codes for an RNA binding protein with novel regulatory domains. , 1994, Oncogene.

[25]  M. Roussel,et al.  DNA-binding and transcriptional activation properties of the EWS-FLI-1 fusion protein resulting from the t(11;22) translocation in Ewing sarcoma , 1994, Molecular and cellular biology.

[26]  Juli D. Klemm,et al.  Crystal structure of the Oct-1 POU domain bound to an octamer site: DNA recognition with tethered DNA-binding modules , 1994, Cell.

[27]  E. Turner,et al.  Brn-3.0: a POU-domain protein expressed in the sensory, immune, and endocrine systems that functions on elements distinct from known octamer motifs. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[28]  C. Verrijzer,et al.  POU domain transcription factors. , 1993, Biochimica et biophysica acta.

[29]  P. Sharp,et al.  Recognition of the surface of a homeo domain protein. , 1992, Genes & development.

[30]  D. Latchman,et al.  A novel POU family transcription factor is closely related to Brn-3 but has a distinct expression pattern in neuronal cells. , 1992, Nucleic acids research.

[31]  S. Bevan,et al.  Novel cell lines display properties of nociceptive sensory neurons , 1990, Proceedings of the Royal Society of London. Series B: Biological Sciences.

[32]  L. Swanson,et al.  Expression of a large family of POU-domain regulatory genes in mammalian brain development , 1989, Nature.

[33]  P. Rigby,et al.  High efficiency gene transfer into mammalian cells. , 1984, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[34]  W. Gerald,et al.  Molecular biology of the Ewing's sarcoma/primitive neuroectodermal tumor family. , 2000, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[35]  D. Latchman,et al.  18 The N-terminal domain unique to the long form of the Brn-3a transcription factor is essential to protect neuronal cells from apoptosis and for the activation of Bcl-2 gene , 1998 .

[36]  D. Latchman,et al.  The POU domain factors Brn-3a and Brn-3b interact with the estrogen receptor and differentially regulate transcriptional activity via an ERE , 1997 .