FBXO21 mediates the ubiquitylation and proteasomal degradation of EID1

Koki Watanabe, Kanae Yumimoto, Keiichi I. Nakayama

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Although identification of substrates for ubiquitin ligase (E3) is important for understanding its biological functions, detection of the interaction between an E3 and its substrates has remained challenging. We recently developed a new approach, termed differential proteomics-based identification of ubiquitylation substrates (DiPIUS), for the discovery of substrates of a given E3 ligase. We have now applied this approach to an uncharacterized human F-box protein, FBXO21, which serves as the substrate-recognition subunit of a SKP1-CUL1-F-box protein (SCF)-type E3, thereby identifying EID1 (EP300-interacting inhibitor of differentiation 1) as a candidate substrate. The central and COOH-terminal portion of FBXO21 was found to interact with the COOH-terminal region of EID1 in transfected cells. Over-expression of FBXO21 resulted in the down-regulation of EID1, whereas disruption of the FBXO21 gene with the CRISPR/Cas9 system stabilized EID1 and led to its accumulation in both the cytoplasm and nucleus. An in vitro ubiquitylation assay showed that EID1 is a direct substrate of SCFFBXO21. Collectively, our results suggest that EID1 is a bona fide substrate of FBXO21 and that the control of EID1 abundance by SCFFBXO21 might affect the transcriptional repression activity of EID1.

Original languageEnglish
Pages (from-to)667-674
Number of pages8
JournalGenes to Cells
Volume20
Issue number8
DOIs
Publication statusPublished - Aug 1 2015

Fingerprint

F-Box Proteins
Ubiquitin-Protein Ligases
Ubiquitination
Clustered Regularly Interspaced Short Palindromic Repeats
Proteomics
Cytoplasm
Down-Regulation
Genes

All Science Journal Classification (ASJC) codes

  • Genetics
  • Cell Biology

Cite this

FBXO21 mediates the ubiquitylation and proteasomal degradation of EID1. / Watanabe, Koki; Yumimoto, Kanae; Nakayama, Keiichi I.

In: Genes to Cells, Vol. 20, No. 8, 01.08.2015, p. 667-674.

Research output: Contribution to journalArticle

Watanabe, Koki ; Yumimoto, Kanae ; Nakayama, Keiichi I. / FBXO21 mediates the ubiquitylation and proteasomal degradation of EID1. In: Genes to Cells. 2015 ; Vol. 20, No. 8. pp. 667-674.
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