Identification of target genes for the Ewing's sarcoma EWS/FLI fusion protein by representational difference analysis.

Abstract

The EWS/FLI-1 fusion gene results from the 11;22 chromosomal translocation in Ewing's sarcoma. The product of the gene is one of a growing number of structurally altered transcription factors implicated in oncogenesis. We have employed a subtractive cloning strategy of representational difference analysis in conjunction with a model transformation system to identify genes transcribed in response to EWS/FLI. We have characterized eight transcripts that are dependent on EWS/FLI for expression and two transcripts that are repressed in response to EWS/FLI. Three of the former were identified by sequence analysis as stromelysin 1, a murine homolog of cytochrome P-450 F1 and cytokeratin 15. Stromelysin 1 is induced rapidly after expression of EWS/FLI, suggesting that the stromelysin 1 gene may be a direct target gene of EWS/FLI. These results demonstrate that expression of EWS/FLI leads to significant changes in the transcription of specific genes and that these effects are at least partially distinct from those caused by expression of germ line FLI-1. The representational difference analysis technique can potentially be applied to investigate transformation pathways activated by a broad array of genes in different tumor systems.

• Authors: B S Braun, R Frieden, S L Lessnick, W A May, C T Denny
• Journal: Molecular and cellular biology (Mol Cell Biol) Vol. 15 Issue 8 Pg. 4623-30 (Aug 1995) ISSN: 0270-7306 [Print] United States
• PMID: 7623854 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• DNA, Complementary
• DNA-Binding Proteins
• Heterogeneous-Nuclear Ribonucleoproteins
• Neoplasm Proteins
• Nuclear Proteins
• Proto-Oncogene Protein c-fli-1
• Proto-Oncogene Proteins
• RNA, Messenger
• RNA, Neoplasm
• RNA-Binding Protein EWS
• RNA-Binding Proteins
• Ribonucleoproteins
• Trans-Activators
• Metalloendopeptidases
• Matrix Metalloproteinase 3

Topics

• Base Sequence
• DNA, Complementary (genetics)
• DNA-Binding Proteins (genetics, metabolism)
• Gene Expression Regulation, Neoplastic
• Heterogeneous-Nuclear Ribonucleoproteins
• Matrix Metalloproteinase 3
• Metalloendopeptidases (genetics)
• Molecular Biology (methods)
• Molecular Sequence Data
• Neoplasm Proteins (genetics, metabolism)
• Nuclear Proteins (genetics, metabolism)
• Proto-Oncogene Protein c-fli-1
• Proto-Oncogene Proteins
• RNA, Messenger (genetics)
• RNA, Neoplasm (genetics)
• RNA-Binding Protein EWS
• RNA-Binding Proteins (genetics, metabolism)
• Ribonucleoproteins (genetics, metabolism)
• Sarcoma, Ewing (genetics)
• Selection, Genetic
• Trans-Activators (genetics, metabolism)
• Up-Regulation