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Chromosomal localization of Ewing sarcoma EWSR1/FLI1 protein promotes the induction of aneuploidy.
| Content Provider | Europe PMC |
|---|---|
| Author | Park, Hyewon Kim, Haeyoung Hassebroek, Victoria Azuma, Yoshiaki Slawson, Chad Azuma, Mizuki |
| Copyright Year | 2020 |
| Abstract | Ewing sarcoma is a pediatric bone cancer that expresses the chimeric protein EWSR1/FLI1. We previously demonstrated that EWSR1/FLI1 impairs the localization of Aurora B kinase to the midzone (the midline structure located between segregating chromosomes) during anaphase. While localization of Aurora B is essential for faithful cell division, it is unknown whether interference with midzone organization by EWSR1/FLI1 induces aneuploidy. To address this, we generated stable Tet-on inducible cell lines with EWSR1/FLI1, using CRISPR/Cas9 technology to integrate the transgene at the safe-harbor AAVS1 locus in DLD-1 cells. Induced cells expressing EWSR1/FLI1 displayed an increased incidence of aberrant localization of Aurora B, and greater levels of aneuploidy, compared with noninduced cells. Furthermore, the expression of EWSR1/FLI1-T79A, containing a threonine (Thr) to alanine (Ala) substitution at amino acid 79, failed to induce these phenotypes, indicating that Thr 79 is critical for EWSR1/FLI1 interference with mitosis. In contrast, the phosphomimetic mutant EWSR1/FLI1-T79D (Thr to aspartic acid (Asp)) retained the high activity as wild-type EWSR1/FLI1. Together, these findings suggest that phosphorylation of EWSR1/FLI1 at Thr 79 promotes the colocalization of EWSR1/FLI1 and Aurora B on the chromosomes during prophase and metaphase and, in addition, impairs the localization of Aurora B during anaphase, leading to induction of aneuploidy. This is the first demonstration of the mechanism for EWSR1/FLI1-dependent induction of aneuploidy associated with mitotic dysfunction and the identification of the phosphorylation of the Thr 79 of EWSR1/FLI1 as a critical residue required for this induction. |
| ISSN | 00219258 |
| Volume Number | 296 |
| PubMed Central reference number | PMC7857440 |
| PubMed reference number | 33293370 |
| Journal | The Journal of Biological Chemistry [J. Biol. Chem] |
| e-ISSN | 1083351X |
| DOI | 10.1074/jbc.RA120.014328 |
| Language | English |
| Publisher | American Society for Biochemistry and Molecular Biology |
| Publisher Date | 2020-12-10 |
| Access Restriction | Open |
| Rights License | This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). © 2020 The Authors |
| Subject Keyword | Ewing sarcoma EWSR1/FLI1 Aurora B aneuploidy mitosis CIN, chromosome instability Dox, doxycycline hMSC, human mesenchymal stem cells HSD, honestly significant difference PTM, posttranslational modification SAC, spindle assembly checkpoint |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Molecular Biology Biochemistry |
