EWS and ATF-1 gene fusion induced by t(12;22) translocation in malignant melanoma of soft parts

Zucman, Jessica; Delattre, Olivier; Desmaze, Chantal; Epstein, Alan L.; Stenman, Göran; Speleman, Frank; Fletchers, Christopher D. M.; Aurias, Alain; Thomas, Gilles

doi:10.1038/ng0893-341

Cited by 464 publications

(315 citation statements)

References 20 publications

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“…For cell viability assays, stable cell transformants were selected by G418 and viable colonies stained with Coomassie Blue (12 days post selection). We first tested two cell lines (Kao and MST1) derived from Soft Tissue Clear Cell Sarcoma (ST-CCS), a member of the Ewing's family of tumors (Zucman et al, 1993;Dim et al, 2007;Wang et al, 2009). Both hsRpb4 and hsRpb7 shRNAs were toxic to Kao (data not shown) and MST1 cells (Fig.…”

Section: Or Cell Lines (Kawl Unpublished Results)mentioning

confidence: 99%

The RNA Pol II sub-complex hsRpb4/7 is required for viability of multiple human cell lines

Zhao¹,

Li²,

Ng³

et al. 2012

Protein Cell

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The evolutionarily conserved RNA Polymerase II Rpb4/7 sub-complex has been thoroughly studied in yeast and impacts gene expression at multiple levels including transcription, mRNA processing and decay. In addition Rpb4/7 exerts differential effects on gene expression in yeast and Rpb4 is not obligatory for yeast (S. cerevisiae) survival. Specialised roles for human (hs) Rpb4/7 have not been extensively described and we have probed this question by depleting hsRpb4/7 in established human cell lines using RNA interference. We find that Rpb4/7 protein levels are inter-dependent and accordingly, the functional effects of depleting either protein are co-incident. hsRpb4/7 exhibits gene-specific effects and cells initially remain viable upon hsRpb4/7 depletion. However prolonged hsRpb4/7 depletion is cytotoxic in the range of cell lines tested. Protracted cell death occurs by an unknown mechanism and in some cases is accompanied by a pronounced elongated cell morphology. In conclusion we provide evidence for a gene-specific role of hsRpb4/7 in human cell viability.

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Section: Or Cell Lines (Kawl Unpublished Results)mentioning

confidence: 99%

The RNA Pol II sub-complex hsRpb4/7 is required for viability of multiple human cell lines

Zhao¹,

Li²,

Ng³

et al. 2012

Protein Cell

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“…In each translocation case involving EWS1, the chimeric product contains the NTD-EWS juxtaposed next to the DNA binding domain of a transcription factor. The fusion partners involve: (i) the ETS-related DNA binding domain of FLI1, ERG, or ETV1 in Ewing's sarcoma and related primitive neuroectodermal family of tumors (Delattre et al, 1992;Sorensen et al, 1994;Jeon et al, 1995), (ii) the bZIP protein dimerization and DNA binding domain of ATF-1 in clear-cell sarcoma of soft tissue (Zucman et al, 1993), and (iii) three of the four WT1 zinc ®ngers in DSRCT (Ladanyi and Gerald, 1994;Gerald et al, 1995).…”

Section: Discussionmentioning

confidence: 99%

The DNA binding domains of the WT1 tumor suppressor gene product and chimeric EWS/WT1 oncoprotein are functionally distinct

1998

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The t(11; 22)(p13; q12) translocation associated with desmosplastic small round cell tumor results in a chimeric molecule fusing the amino terminal domain (NTD) of the EWS1 gene to three of the four carboxyterminal zinc ®ngers of the WT1 tumor suppressor gene. Since the DNA binding domains of WT1 and EWS/WT1 are structurally di erent, we have assessed the functional consequences of the EWS/WT1 fusion. We ®nd that the EWS/WT1 protein has a higher binding a nity for a given recognition target than the WT1 product. This is unlike other fusion products involving translocation of the NTD of EWS to DNA binding domains in which DNA binding speci®city and a nity is not changed. We demonstrate that EWS/WT1 is a nuclear protein and that the NTD of EWS contains (a) nuclear localization signal(s). We also ®nd that the integrity of a domain within the WT1 zinc ®ngers, responsible for mediating interaction between WT1 and the transcriptional repressor par-4, is disrupted in the EWS/WT1 fusion product. Deletion analysis of the NTD of EWS indicated that integrity of the entire domain was necessary to achieve full transactivation potential.

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“…Fusions of EWS to other families of transcription factors have been observed in di erent types of tumors, including CHOP in myxoid liposarcoma (Panagopoulos et al, 1996), ATF-1 in malignant melanoma of soft parts (Zucman et al, 1993a;Fujimura et al, 1996), WT1 in intra-abdominal desmoplastic small round cell tumor (Ladanyi and Gerald, 1994) and to orphan nuclear receptors in myxoid chondrosarcoma (Labelle et al, 1995;Clark et al, 1996). TLS/FUS-CHOP and TLS/FUS-ERG chimeras have been described in myxoid liposarcoma (Rabbitts et al, 1993;Crozat et al, 1993) and acute myeloid leukemia (Panagopoulos et al, 1994;Ichikawa et al, 1994), respectively.…”

Section: Introductionmentioning

confidence: 99%

Oncogenic EWS-Fli1 interacts with hsRPB7, a subunit of human RNA polymerase II

et al. 1998

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As a result of the t(11;22)(q24;q12) chromosomal translocation characterizing the Ewing family of tumors (ET), the amino terminal portion of EWS, an RNA binding protein of unknown function, is fused to the DNA-binding domain of the ets transcription factor Fli1. The hybrid EWS-Fli1 protein acts as a strong transcriptional activator and, in contrast to wildtype Fli1, is a potent transforming agent. Similar rearrangements involving EWS or the highly homologous TLS with various transcription factors have been found in several types of human tumors. Employing yeast twohybrid cloning we isolated the seventh largest subunit of human RNA polymerase II (hsRPB7) as a protein that speci®cally interacts with the amino terminus of EWS. This association was con®rmed by in vitro immunocoprecipitation. In nuclear extracts, hsRPB7 was found to copurify with EWS-Fli1 but not with Fli1. Overexpression of recombinant hsRPB7 speci®cally increased gene activation by EWS-chimeric transcription factors. Replacement of the EWS portion by hsRPB7 in the oncogenic fusion protein restored the transactivating potential of the chimera. Our results suggest that the interaction of the amino terminus of EWS with hsRPB7 contributes to the transactivation function of EWS-Fli1 and, since hsRPB7 has characteristics of a regulatory subunit of RNA polymerase II, may in¯uence promoter selectivity.

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EWS and ATF-1 gene fusion induced by t(12;22) translocation in malignant melanoma of soft parts

Cited by 464 publications

References 20 publications

The RNA Pol II sub-complex hsRpb4/7 is required for viability of multiple human cell lines

The RNA Pol II sub-complex hsRpb4/7 is required for viability of multiple human cell lines

The DNA binding domains of the WT1 tumor suppressor gene product and chimeric EWS/WT1 oncoprotein are functionally distinct

Oncogenic EWS-Fli1 interacts with hsRPB7, a subunit of human RNA polymerase II

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