SCF Fbxw7 ubiquitylates KLF7 for degradation in a manner dependent on GSK-3-mediated phosphorylation

Shigeaki Sugiyama, Kanae Yumimoto, Ippei Inoue, Keiichi Nakayama

研究成果: ジャーナルへの寄稿記事

抄録

The biological relation between ubiquitin ligases and their substrates has been largely unclear. We previously developed a method—differential proteomics-based identification of ubiquitylation substrates (DiPIUS)—for the comprehensive identification of substrates for a given ubiquitin ligase. We have now applied DiPIUS to the F-box protein Fbxw7 in three cell lines (mHepa, Neuro2A and C2C12) and thereby identified Krüppel-like factor 7 (KLF7) as a candidate substrate of the SCF Fbxw7 ubiquitin ligase complex. KLF7 was shown to interact with Fbxw7 and to undergo Fbxw7-mediated polyubiquitylation. The stability of KLF7 was increased by depletion of Fbxw7, mutation of a putative Cdc4 phosphodegron (CPD) of KLF7 or exposure to inhibitors of glycogen synthase kinase-3 (GSK-3). Over-expression of Fbxw7 in Neuro2A cells down-regulated expression of the p21 Cip1 gene, which is a transcriptional target of KLF7 in neuronal differentiation and maintenance. Despite the presence of an almost identical CPD sequence in KLF6, the closest paralog of KLF7, mutation of this sequence affected neither the interaction of KLF6 with Fbxw7 nor its half-life. Our results suggest that KLF7, but not KLF6, is a bona fide substrate of SCF Fbxw7 , and that control of KLF7 abundance by SCF Fbxw7 might contribute to the regulation of neuronal differentiation and maintenance.

元の言語英語
ページ(範囲)354-365
ページ数12
ジャーナルGenes to Cells
24
発行部数5
DOI
出版物ステータス出版済み - 5 1 2019

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Glycogen Synthase Kinase 3
Ligases
Ubiquitin
SKP Cullin F-Box Protein Ligases
Maintenance
Phosphorylation
F-Box Proteins
Mutation
Ubiquitination
Proteomics
Half-Life
Cell Line
Genes

All Science Journal Classification (ASJC) codes

  • Genetics
  • Cell Biology

これを引用

SCF Fbxw7 ubiquitylates KLF7 for degradation in a manner dependent on GSK-3-mediated phosphorylation . / Sugiyama, Shigeaki; Yumimoto, Kanae; Inoue, Ippei; Nakayama, Keiichi.

:: Genes to Cells, 巻 24, 番号 5, 01.05.2019, p. 354-365.

研究成果: ジャーナルへの寄稿記事

Sugiyama, Shigeaki ; Yumimoto, Kanae ; Inoue, Ippei ; Nakayama, Keiichi. / SCF Fbxw7 ubiquitylates KLF7 for degradation in a manner dependent on GSK-3-mediated phosphorylation :: Genes to Cells. 2019 ; 巻 24, 番号 5. pp. 354-365.
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AB - The biological relation between ubiquitin ligases and their substrates has been largely unclear. We previously developed a method—differential proteomics-based identification of ubiquitylation substrates (DiPIUS)—for the comprehensive identification of substrates for a given ubiquitin ligase. We have now applied DiPIUS to the F-box protein Fbxw7 in three cell lines (mHepa, Neuro2A and C2C12) and thereby identified Krüppel-like factor 7 (KLF7) as a candidate substrate of the SCF Fbxw7 ubiquitin ligase complex. KLF7 was shown to interact with Fbxw7 and to undergo Fbxw7-mediated polyubiquitylation. The stability of KLF7 was increased by depletion of Fbxw7, mutation of a putative Cdc4 phosphodegron (CPD) of KLF7 or exposure to inhibitors of glycogen synthase kinase-3 (GSK-3). Over-expression of Fbxw7 in Neuro2A cells down-regulated expression of the p21 Cip1 gene, which is a transcriptional target of KLF7 in neuronal differentiation and maintenance. Despite the presence of an almost identical CPD sequence in KLF6, the closest paralog of KLF7, mutation of this sequence affected neither the interaction of KLF6 with Fbxw7 nor its half-life. Our results suggest that KLF7, but not KLF6, is a bona fide substrate of SCF Fbxw7 , and that control of KLF7 abundance by SCF Fbxw7 might contribute to the regulation of neuronal differentiation and maintenance.

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