The cooh-terminal domain of FLI-1 is necessary for full tumorigenesis and transcriptional modulation by EWS/FLI-11,2

Afsane Arvand, Michael A. Teitell, Scott Welford, Michael A. Teitell, Christopher T. Denny, Michael A. Teitell, Christopher T. Denny, Christopher T. Denny

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

More than 85 % of Ewing's family tumors carry a specific chromosomal translocation that fuses the NH2 terminus of the EWS gene to the COOH terminus of the FLI1 transcription factor. It has been shown previously that both the transactivation domain encoded by EWS and the DNA binding domain of FLI1 were necessary for transforming cells to anchorage independence. We now report that a COOH-terminal domain in addition to the FLI1 DNA binding domain is necessary to promote cellular transformation. NIH 3T3 cells expressing a COOH-terminal deletion mutant (EWS/FLI1 ΔC) have a greatly reduced capability to form colonies in soft agar and tumors in severe combined immunodeficient mice. The rate of tumor formation for NIH 3T3 that express EWS/FLI1 ΔC is 50 days, whereas EWS/FLI1 forms tumors within 22 days. In addition, cells expressing the EWS/FLI1 ΔC mutant failed to completely demonstrate the round-cell histology that is seen in both Ewing's tumor cell lines and NIH 3T3 cells expressing full-length EWS/FLI1. Northern and microarray analyses were performed to assess the effect of loss of the FLI1 COOH terminus on transcriptional modulation of EWS/FLI1 target genes. We found that although EWS/FLI1 ΔC up-regulates smaller numbers of genes (21 genes) compared with EWS/FLI1 (34 genes), 41% of the EWS/FLI1 targets were also up-regulated by EWS/FLI1 ΔC. On the other hand, EWS/FLI1 ΔC is unable to down-regulate genes (3 genes) as efficiently as EWS/FLI1 (39 genes) with only one target gene repressed by both fusion constructs. Our study indicates that the EWS/FLI1 transcription factor has strong transcriptional activating as well as repressing properties and suggests that transcriptional activation and repression of target genes may occur through biochemically different mechanisms.

Original languageEnglish (US)
Pages (from-to)5311-5317
Number of pages7
JournalCancer Research
Volume61
Issue number13
StatePublished - Jul 1 2001
Externally publishedYes

Fingerprint

Carcinogenesis
Genes
NIH 3T3 Cells
Ewing's Sarcoma
EWS-FLI fusion protein
Transcriptional Activation
Transcription Factors
Neoplasms
Genetic Translocation
SCID Mice
DNA
Microarray Analysis
Tumor Cell Line
Agar
Histology
Up-Regulation
Down-Regulation

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Arvand, A., Teitell, M. A., Welford, S., Teitell, M. A., Denny, C. T., Teitell, M. A., ... Denny, C. T. (2001). The cooh-terminal domain of FLI-1 is necessary for full tumorigenesis and transcriptional modulation by EWS/FLI-11,2 Cancer Research, 61(13), 5311-5317.

The cooh-terminal domain of FLI-1 is necessary for full tumorigenesis and transcriptional modulation by EWS/FLI-11,2 . / Arvand, Afsane; Teitell, Michael A.; Welford, Scott; Teitell, Michael A.; Denny, Christopher T.; Teitell, Michael A.; Denny, Christopher T.; Denny, Christopher T.

In: Cancer Research, Vol. 61, No. 13, 01.07.2001, p. 5311-5317.

Research output: Contribution to journalArticle

Arvand, A, Teitell, MA, Welford, S, Teitell, MA, Denny, CT, Teitell, MA, Denny, CT & Denny, CT 2001, 'The cooh-terminal domain of FLI-1 is necessary for full tumorigenesis and transcriptional modulation by EWS/FLI-11,2 ', Cancer Research, vol. 61, no. 13, pp. 5311-5317.
Arvand, Afsane ; Teitell, Michael A. ; Welford, Scott ; Teitell, Michael A. ; Denny, Christopher T. ; Teitell, Michael A. ; Denny, Christopher T. ; Denny, Christopher T. / The cooh-terminal domain of FLI-1 is necessary for full tumorigenesis and transcriptional modulation by EWS/FLI-11,2 In: Cancer Research. 2001 ; Vol. 61, No. 13. pp. 5311-5317.
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abstract = "More than 85 {\%} of Ewing's family tumors carry a specific chromosomal translocation that fuses the NH2 terminus of the EWS gene to the COOH terminus of the FLI1 transcription factor. It has been shown previously that both the transactivation domain encoded by EWS and the DNA binding domain of FLI1 were necessary for transforming cells to anchorage independence. We now report that a COOH-terminal domain in addition to the FLI1 DNA binding domain is necessary to promote cellular transformation. NIH 3T3 cells expressing a COOH-terminal deletion mutant (EWS/FLI1 ΔC) have a greatly reduced capability to form colonies in soft agar and tumors in severe combined immunodeficient mice. The rate of tumor formation for NIH 3T3 that express EWS/FLI1 ΔC is 50 days, whereas EWS/FLI1 forms tumors within 22 days. In addition, cells expressing the EWS/FLI1 ΔC mutant failed to completely demonstrate the round-cell histology that is seen in both Ewing's tumor cell lines and NIH 3T3 cells expressing full-length EWS/FLI1. Northern and microarray analyses were performed to assess the effect of loss of the FLI1 COOH terminus on transcriptional modulation of EWS/FLI1 target genes. We found that although EWS/FLI1 ΔC up-regulates smaller numbers of genes (21 genes) compared with EWS/FLI1 (34 genes), 41{\%} of the EWS/FLI1 targets were also up-regulated by EWS/FLI1 ΔC. On the other hand, EWS/FLI1 ΔC is unable to down-regulate genes (3 genes) as efficiently as EWS/FLI1 (39 genes) with only one target gene repressed by both fusion constructs. Our study indicates that the EWS/FLI1 transcription factor has strong transcriptional activating as well as repressing properties and suggests that transcriptional activation and repression of target genes may occur through biochemically different mechanisms.",
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