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Ahmed NS, Harrell LM, Wieland DR, et al. Fusion protein EWS-FLI1 is incorporated into a protein granule in cells. RNA. 2021;doi: 10.1261/rna.078827.121.
Ahmed, N. S., Harrell, L. M., Wieland, D. R., Lay, M. A., Thompson, V. F., & Schwartz, J. C. (2021). Fusion protein EWS-FLI1 is incorporated into a protein granule in cells. RNA (New York, N.Y.), . https://doi.org/10.1261/rna.078827.121
Ahmed, Nasiha S, et al. "Fusion protein EWS-FLI1 is incorporated into a protein granule in cells." RNA (New York, N.Y.) vol. (2021). doi: https://doi.org/10.1261/rna.078827.121
Ahmed NS, Harrell LM, Wieland DR, Lay MA, Thompson VF, Schwartz JC. Fusion protein EWS-FLI1 is incorporated into a protein granule in cells. RNA. 2021 May 25; doi: 10.1261/rna.078827.121. Epub 2021 May 25. PMID: 34035145; PMCID: PMC8284321.
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RNA. 2021 May 25; doi: 10.1261/rna.078827.121. Epub 2021 May 25.

Fusion protein EWS-FLI1 is incorporated into a protein granule in cells.

RNA (New York, N.Y.)

Nasiha S Ahmed, Lucas M Harrell, Daniel R Wieland, Michelle A Lay, Valery F Thompson, Jacob C Schwartz

Affiliations

  1. Department of Molecular and Cellular Biology, The University of Arizona, Tucson, AZ 85719.
  2. Department of Chemistry and Biochemistry, The University of Arizona, Tucson, AZ 85719.
  3. Department of Chemistry and Biochemistry, The University of Arizona, Tucson, AZ 85719 [email protected].

PMID: 34035145 PMCID: PMC8284321 DOI: 10.1261/rna.078827.121

Abstract

Ewing sarcoma is driven by fusion proteins containing a low complexity (LC) domain that is intrinsically disordered and a powerful transcriptional regulator. The most common fusion protein found in Ewing sarcoma, EWS-FLI1, takes its LC domain from the RNA-binding protein EWSR1 (Ewing Sarcoma RNA-binding protein 1) and a DNA-binding domain from the transcription factor FLI1 (Friend Leukemia Virus Integration 1). EWS-FLI1 can bind RNA polymerase II (RNA Pol II) and self-assemble through its low-complexity (LC) domain. The ability of RNA-binding proteins like EWSR1 to self-assemble or phase separate in cells has raised questions about the contribution of this process to EWS-FLI1 activity. We examined EWSR1 and EWS-FLI1 activity in Ewing sarcoma cells by siRNA-mediated knockdown and RNA-seq analysis. More transcripts were affected by the EWSR1 knockdown than expected and these included many EWS-FLI1 regulated genes. We reevaluated physical interactions between EWS-FLI1, EWSR1, and RNA Pol II, and employed a cross-linking based strategy to investigate protein assemblies associated with the proteins. The LC domain of EWS-FLI1 was required for the assemblies observed to form in cells. These results offer new insights into a protein assembly that may enable EWS-FLI1 to bind its wide network of protein partners and contribute to regulation of gene expression in Ewing sarcoma.

Published by Cold Spring Harbor Laboratory Press for the RNA Society.

Keywords: Ewing sarcoma; fusion proteins; granules; phase separation; transcription

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