Abstract | BACKGROUND: METHODS: In-silico model of EWS-FLI1 fusion protein was analysed for ligand binding sites, and a putative region ( amino acid (aa) 251-343 of the type 1 fusion protein) in the vicinity of the fusion junction was cloned and expressed using bacterial expression. The recombinant protein was characterized by Circular Dichroism (CD). We then expressed aa 251-280 ectopically in Ewing's sarcoma cell-line and its effect on cell proliferation, tumorigenicity and expression of EWS-FLI1 target genes were analysed. RESULTS: Our modelling analysis indicated that Junction region (aa 251-343) encompasses potential ligand biding sites in the EWS-FLI1 protein and when expressed in bacteria was present as soluble form. Ectopically expressing this region in Ewing's sarcoma cells inhibited tumorigenicity, and EWS-FLI1 target genes indicating a dominant negative biological effect. CONCLUSIONS:
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Authors | Babu Jully, Ramshankar Vijayalakshmi, Gopisetty Gopal, Kesavan Sabitha, Thangarajan Rajkumar |
Journal | BMC cancer
(BMC Cancer)
Vol. 12
Pg. 513
(Nov 12 2012)
ISSN: 1471-2407 [Electronic] England |
PMID | 23145994
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- EWS-FLI fusion protein
- Homeobox Protein Nkx-2.2
- Homeodomain Proteins
- MYC protein, human
- Oncogene Proteins, Fusion
- Peptides
- Proto-Oncogene Protein c-fli-1
- Proto-Oncogene Proteins c-myc
- RNA-Binding Protein EWS
- Recombinant Proteins
- Transcription Factors
- Zebrafish Proteins
- Cyclin D1
- EZH2 protein, human
- Enhancer of Zeste Homolog 2 Protein
- Polycomb Repressive Complex 2
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Topics |
- Amino Acid Sequence
- Cell Line, Tumor
- Cell Proliferation
- Circular Dichroism
- Cyclin D1
(genetics, metabolism)
- Enhancer of Zeste Homolog 2 Protein
- Epithelial-Mesenchymal Transition
(genetics)
- Gene Expression Regulation, Neoplastic
- Homeobox Protein Nkx-2.2
- Homeodomain Proteins
(genetics, metabolism)
- Humans
- Immunoblotting
- MCF-7 Cells
- Molecular Sequence Data
- Oncogene Proteins, Fusion
(chemistry, genetics, metabolism)
- Peptides
(genetics, metabolism)
- Polycomb Repressive Complex 2
(genetics, metabolism)
- Proto-Oncogene Protein c-fli-1
(chemistry, genetics, metabolism)
- Proto-Oncogene Proteins c-myc
(genetics, metabolism)
- RNA-Binding Protein EWS
(chemistry, genetics, metabolism)
- Recombinant Proteins
(chemistry, metabolism)
- Reverse Transcriptase Polymerase Chain Reaction
- Sarcoma, Ewing
(genetics, metabolism, pathology)
- Transcription Factors
(genetics, metabolism)
- Transfection
- Zebrafish Proteins
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