RRC ID 36791
著者 Nakajima T, Kitagawa K, Ohhata T, Sakai S, Uchida C, Shibata K, Minegishi N, Yumimoto K, Nakayama KI, Masumoto K, Katou F, Niida H, Kitagawa M.
タイトル Regulation of GATA-binding protein 2 levels via ubiquitin-dependent degradation by Fbw7: involvement of cyclin B-cyclin-dependent kinase 1-mediated phosphorylation of THR176 in GATA-binding protein 2.
ジャーナル J Biol Chem
Abstract A GATA family transcription factor, GATA-binding protein 2 (GATA2), participates in cell growth and differentiation of various cells, such as hematopoietic stem cells. Although its expression level is controlled by transcriptional induction and proteolytic degradation, the responsible E3 ligase has not been identified. Here, we demonstrate that F-box/WD repeat-containing protein 7 (Fbw7/Fbxw7), a component of Skp1, Cullin 1, F-box-containing complex (SCF)-type E3 ligase, is an E3 ligase for GATA2. GATA2 contains a cell division control protein 4 (Cdc4) phosphodegron (CPD), a consensus motif for ubiquitylation by Fbw7, which includes Thr(176). Ectopic expression of Fbw7 destabilized GATA2 and promoted its proteasomal degradation. Substitution of threonine 176 to alanine in GATA2 inhibited binding with Fbw7, and the ubiquitylation and degradation of GATA2 by Fbw7 was suppressed. The CPD kinase, which mediates the phosphorylation of Thr(176), was cyclin B-cyclin-dependent kinase 1 (CDK1). Moreover, depletion of endogenous Fbw7 stabilized endogenous GATA2 in K562 cells. Conditional Fbw7 depletion in mice increased GATA2 levels in hematopoietic stem cells and myeloid progenitors at the early stage. Increased GATA2 levels in Fbw7-conditional knock-out mice were correlated with a decrease in a c-Kit high expressing population of myeloid progenitor cells. Our results suggest that Fbw7 is a bona fide E3 ubiquitin ligase for GATA2 in vivo.
巻・号 290(16)
ページ 10368-81
公開日 2015-4-17
DOI 10.1074/jbc.M114.613018
PII S0021-9258(20)42683-3
PMID 25670854
PMC PMC4400347
MeSH Amino Acid Motifs Animals CDC2 Protein Kinase Cell Cycle Proteins / antagonists & inhibitors Cell Cycle Proteins / genetics* Cell Cycle Proteins / metabolism Cell Differentiation Cyclin B / genetics* Cyclin B / metabolism Cyclin-Dependent Kinases / genetics* Cyclin-Dependent Kinases / metabolism F-Box Proteins / antagonists & inhibitors F-Box Proteins / genetics* F-Box Proteins / metabolism F-Box-WD Repeat-Containing Protein 7 GATA2 Transcription Factor / antagonists & inhibitors GATA2 Transcription Factor / genetics* GATA2 Transcription Factor / metabolism Gene Expression Regulation Glycogen Synthase Kinase 3 / genetics Glycogen Synthase Kinase 3 / metabolism HEK293 Cells HeLa Cells Humans K562 Cells Mice Mice, Knockout Molecular Sequence Data Mutation Phosphorylation Proteasome Endopeptidase Complex / metabolism Protein Binding Proteolysis RNA, Small Interfering / genetics RNA, Small Interfering / metabolism Signal Transduction Ubiquitin / genetics Ubiquitin / metabolism Ubiquitin-Protein Ligases / antagonists & inhibitors Ubiquitin-Protein Ligases / genetics* Ubiquitin-Protein Ligases / metabolism Ubiquitination
IF 4.238
引用数 13
WOS 分野 BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
ヒト・動物細胞 K562