The Transcriptional Repressor ZFM1 Interacts with and Modulates the Ability of EWS to Activate Transcription*

The ZFM1 protein is both a transcriptional repressor and identical to the splicing factor SF1. ZFM1 was shown to interact with and repress transcription from the glycine, glutamine, serine, and threonine-rich transcription activation domain of the sea urchin transcription factor, stage-specific activator protein (SSAP). EWS, a human protein involved in cellular transformation in Ewing’s sarcoma tumors, contains an NH2-terminal transcriptional activation domain (NTD) which resembles that of SSAP in both amino acid composition and the ability to drive transcription to levels higher than VP16 in most cell types. Here we report that ZFM1 also interacts with EWS in both two-hybrid assays and glutathioneS-transferase pull-down experiments. The region on EWS which interacts with ZFM1 maps to 37 amino acids within its NTD. Overexpression of ZFM1 in HepG2 cells represses the transactivation of reporter gene expression driven by Gal4-EWS-NTD fusion protein and this repression correlates with ZFM1 binding to EWS. Furthermore, two proteins, TLS and hTAFII68, which have extensive homology to EWS, also interact with ZFM1. Recently, it was discovered that EWS/TLS/hTAFII68 are each present in distinct TFIID populations and EWS and hTAFII68 were also found to be associated with the RNA polymerase II holoenzyme. The association of ZFM1 with these proteins implies that one normal cellular function for ZFM1 may be to negatively modulate transcription of target genes coordinated by these cofactors.

[1]  G. Childs,et al.  Human ZFM1 Protein Is a Transcriptional Repressor That Interacts with the Transcription Activation Domain of Stage-specific Activator Protein* , 1998, The Journal of Biological Chemistry.

[2]  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.

[3]  D. Immanuel,et al.  TLS (FUS) binds RNA in vivo and engages in nucleo-cytoplasmic shuttling. , 1997, Journal of cell science.

[4]  Marcienne M Wright,et al.  EWS/FLI1-induced manic fringe renders NIH 3T3 cells tumorigenic , 1997, Nature Genetics.

[5]  M. Rosbash,et al.  The Splicing Factor BBP Interacts Specifically with the Pre-mRNA Branchpoint Sequence UACUAAC , 1997, Cell.

[6]  M. Rosbash,et al.  Cross-Intron Bridging Interactions in the Yeast Commitment Complex Are Conserved in Mammals , 1997, Cell.

[7]  C. Denny,et al.  Biology of EWS/FLI and related fusion genes in Ewing's sarcoma and primitive neuroectodermal tumor. , 1997, Current topics in microbiology and immunology.

[8]  C. Denny,et al.  EAT-2 is a novel SH2 domain containing protein that is up regulated by Ewing's sarcoma EWS/FLI1 fusion gene. , 1996, Oncogene.

[9]  Robert Tjian,et al.  Human TAFII105 Is a Cell Type–Specific TFIID Subunit Related to hTAFII130 , 1996, Cell.

[10]  P. Chambon,et al.  hTAF(II)68, a novel RNA/ssDNA‐binding protein with homology to the pro‐oncoproteins TLS/FUS and EWS is associated with both TFIID and RNA polymerase II. , 1996, The EMBO journal.

[11]  A. Krämer,et al.  Mammalian splicing factor SF1 is encoded by variant cDNAs and binds to RNA. , 1996, RNA.

[12]  J. Defalco,et al.  The embryonic transcription factor stage specific activator protein contains a potent bipartite activation domain that interacts with several RNA polymerase II basal transcription factors. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[13]  D. Israeli,et al.  Isolation of 10 differentially expressed cDNAs in p53-induced apoptosis: activation of the vertebrate homologue of the drosophila seven in absentia gene. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[14]  M. Mann,et al.  Identification of hnRNP P2 as TLS/FUS using electrospray mass spectrometry. , 1995, RNA.

[15]  C. Denny,et al.  Identification of target genes for the Ewing's sarcoma EWS/FLI fusion protein by representational difference analysis , 1995, Molecular and cellular biology.

[16]  J. Defalco,et al.  The embryonic enhancer-binding protein SSAP contains a novel DNA-binding domain which has homology to several RNA-binding proteins , 1995, Molecular and cellular biology.

[17]  C. Denny,et al.  Multiple domains mediate transformation by the Ewing's sarcoma EWS/FLI-1 fusion gene. , 1995, Oncogene.

[18]  T. Rabbitts,et al.  Chromosomal translocations in human cancer , 1994, Nature.

[19]  X. Jacq,et al.  Human TAFII30 is present in a distinct TFIID complex and is required for transcriptional activation by the estrogen receptor , 1994, Cell.

[20]  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.

[21]  T. Toda,et al.  Isolation and characterization of a novel gene encoding nuclear protein at a locus (D11S636) tightly linked to multiple endocrine neoplasia type 1 (MEN1). , 1994, Human molecular genetics.

[22]  C. Denny,et al.  The Ewing's sarcoma EWS/FLI-1 fusion gene encodes a more potent transcriptional activator and is a more powerful transforming gene than FLI-1 , 1993, Molecular and cellular biology.

[23]  N. Mandahl,et al.  Fusion of CHOP to a novel RNA-binding protein in human myxoid liposarcoma , 1993, Nature.

[24]  J. Defalco,et al.  Purification and characterization of the stage-specific embryonic enhancer-binding protein SSAP-1 , 1993, Molecular and cellular biology.

[25]  P. Chambon,et al.  Distinct TFIID complexes mediate the effect of different transcriptional activators. , 1993, The EMBO journal.

[26]  A. Krämer,et al.  Purification of splicing factor SF1, a heat-stable protein that functions in the assembly of a presplicing complex , 1992, Molecular and cellular biology.

[27]  G. Thomas,et al.  Gene fusion with an ETS DNA-binding domain caused by chromosome translocation in human tumours , 1992, Nature.