ELK-1 ubiquitination status and transcriptional activity are modulated independently of F-Box protein FBXO25

Quintero-Barceinas, Reyna Sara; Gehringer, Franziska; Ducker, Charles; Saxton, Janice; Shaw, Peter E.

Abstract

The mitogen-responsive, ETS-domain transcription factor ELK-1 stimulates the expression of immediate early genes at the onset of the cell cycle and participates in early developmental programming. ELK-1 is subject to multiple levels of posttranslational control, including phosphorylation, SUMOylation, and ubiquitination. Recently, removal of monoubiquitin from the ELK-1 ETS domain by the Ubiquitin Specific Protease USP17 was shown to augment ELK-1 transcriptional activity and promote cell proliferation. Here we have used coimmunoprecipitation experiments, protein turnover and ubiquitination assays, RNA-interference and gene expression analyses to examine the possibility that USP17 acts antagonistically with the F-box protein FBXO25, an E3 ubiquitin ligase previously shown to promote ELK-1 ubiquitination and degradation. Our data confirm that FBXO25 and ELK-1 interact in HEK293T cells and that FBXO25 is active toward Hand1 and HAX1, two of its other candidate substrates. However, our data indicate that FBXO25 neither promotes ubiquitination of ELK-1 nor impacts on its transcriptional activity and suggest that an E3 ubiquitin ligase other than FBXO25 regulates ELK-1 ubiquitination and function.

Citation

Quintero-Barceinas, R. S., Gehringer, F., Ducker, C., Saxton, J., & Shaw, P. E. (2021). ELK-1 ubiquitination status and transcriptional activity are modulated independently of F-Box protein FBXO25. Journal of Biological Chemistry, 296, Article 100214. https://doi.org/10.1074/jbc.ra120.014616

• Journal Article Type: Article
• Acceptance Date: Dec 11, 2020
• Online Publication Date: Jan 4, 2021
• Publication Date: 2021
• Deposit Date: Feb 4, 2021
• Publicly Available Date: Feb 4, 2021
• Journal: Journal of Biological Chemistry
• Print ISSN: 0021-9258
• Publisher: American Society for Biochemistry and Molecular Biology
• Peer Reviewed: Peer Reviewed
• Volume: 296
• Article Number: 100214
• DOI: https://doi.org/10.1074/jbc.ra120.014616
• Keywords: gene regulation; ETS transcription factor family; ubiquitin ligase; cell proliferation; development; ubiquitin-specific protease 17 (USP17)
• Public URL: https://nottingham-repository.worktribe.com/output/5291909
• Publisher URL: https://www.sciencedirect.com/science/article/pii/S0021925820002100
• Additional Information: This article is maintained by: Elsevier; Article Title: ELK-1 ubiquitination status and transcriptional activity are modulated independently of F-Box protein FBXO25; Journal Title: Journal of Biological Chemistry; CrossRef DOI link to publisher maintained version: https://doi.org/10.1074/jbc.RA120.014616; Content Type: article; Copyright: © 2020 The Authors. Published by Elsevier Inc on behalf of American Society for Biochemistry and Molecular Biology.