dc.contributor.author | Park, Hyewon | |
dc.contributor.author | Kim, Haeyoung | |
dc.contributor.author | Hassebroek, Victoria | |
dc.contributor.author | Azuma, Yoshiaki | |
dc.contributor.author | Slawson, Chad | |
dc.contributor.author | Azuma, Mizuki | |
dc.date.accessioned | 2022-01-06T21:07:00Z | |
dc.date.available | 2022-01-06T21:07:00Z | |
dc.date.issued | 2020-12-10 | |
dc.identifier.citation | Park, H., Kim, H., Hassebroek, V., Azuma, Y., Slawson, C., & Azuma, M. (2021). Chromosomal localization of Ewing sarcoma EWSR1/FLI1 protein promotes the induction of aneuploidy. The Journal of biological chemistry, 296, 100164. https://doi.org/10.1074/jbc.RA120.014328 | en_US |
dc.identifier.uri | http://hdl.handle.net/1808/32362 | |
dc.description.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. | en_US |
dc.publisher | Elsevier | en_US |
dc.rights | © 2020 THE AUTHORS. Published by Elsevier Inc on behalf of American Society for Biochemistry and Molecular Biology. This is an open access article under the CC-BY license. | en_US |
dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | en_US |
dc.subject | Ewing sarcoma | en_US |
dc.subject | EWSR1/FLI1 | en_US |
dc.subject | Aurora B | en_US |
dc.subject | Aneuploidy | en_US |
dc.subject | Mitosis | en_US |
dc.title | Chromosomal localization of Ewing sarcoma EWSR1/FLI1 protein promotes the induction of aneuploidy | en_US |
dc.type | Article | en_US |
kusw.kuauthor | Park, Hyewon | |
kusw.kuauthor | Kim, Haeyoung | |
kusw.kuauthor | Hassebroek, Victoria | |
kusw.kuauthor | Azuma, Yoshiaki | |
kusw.kuauthor | Azuma, Mizuki | |
kusw.kudepartment | Molecular Biosciences | en_US |
dc.identifier.doi | 10.1074/jbc.RA120.014328 | en_US |
dc.identifier.orcid | https://orcid.org/ 0000-0001-9411-9130 | en_US |
kusw.oaversion | Scholarly/refereed, publisher version | en_US |
kusw.oapolicy | This item meets KU Open Access policy criteria. | en_US |
dc.identifier.pmid | PMC7857440 | en_US |
dc.rights.accessrights | openAccess | en_US |