The E3 Ubiquitin Ligase SMAD Ubiquitination Regulatory Factor 2 Negatively Regulates Krüppel-like Factor 5 Protein

Du, James X.; Hagos, Engda G.; Nandan, Mandayam O.; Bialkowska, Agnieszka B.; Yu, Bing; Yang, Vincent W.

doi:10.1074/jbc.m111.258707

Cited by 44 publications

(41 citation statements)

References 32 publications

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“…A previous study showed that the E3 ubiquitin ligase WWP1 inhibits KLF2 transcriptional activity and targets KLF2 for ubiquitination and degradation in a ubiquitin ligase activity independent manner [23]. The E3 ubiquitin ligase Smurf1can target KLF2 for degradation in H1299 cells [29]. Our data showed that KLF2 protein expression levels increased when HRECs and HUVECs were infected with AX2R lentivirus.…”

Section: Discussionsupporting

confidence: 54%

Overexpression of Annexin A2 Receptor Inhibits Neovascularization via the Promotion of Krüppel-Like Transcription Factor 2

Guo

Song

Zhao

et al. 2018

Cell Physiol Biochem

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Background/Aims: Annexin A2 receptor (AX2R) can mediate annexin A2 signalling and induce apoptosis in a variety of cells, but its role in neovascularization (NV) remains unclear. Krüppel-like transcription factor 2 (KLF2) is known to be expressed in a range of cell types and to participate in a number of processes during development and disease, such as endothelial homeostasis, vasoregulation and vascular growth/remodelling. The aim of our study was to investigate the role of AX2R in NV and the plausible molecular mechanism. Methods: We constructed a eukaryotic overexpression plasmid for AX2R (Lenti-AX2R) by using polymerase chain reaction (PCR). The full-length human AX2R gene was transfected into human retinal endothelial cells (HRECs) and human umbilical vein endothelial cells (HUVECs) using lentivirus vectors to overexpress AX2R. All experiments were divided into three groups: control, negative control (Lenti-EGFP), and Lenti-AX2R.Cell proliferation, cell migration, tube formation, mouse aortic ring assays and mouse matrigel plug assay were applied to analyse the effect of AX2R in NV. Furthermore, we conducted flow cytometry to evaluate whether AX2R could influence the cell cycle. A series of cell cycle-related proteins including cyclin A1, cyclin B1, cyclin D1, cyclin E1, CDK1, and p-CDC2 were detected by WB. The mRNA and protein levels of KLF2, vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor 2 (VEGFR2) were further quantified by RT-PCR and WB to reveal the possible mechanism. Results: Overexpression of AX2R significantly inhibited cell proliferation, migration and tube formation in both types of endothelial cells (ECs), HRECs and HUVECs. It also suppressed vessel sprouting in the mouse aortic ring assay and NV in mouse matrigel plug assay. Furthermore, infection with Lenti-AX2R lentivirus arrested the cell cycle in S/G2 and influenced the expression of a series of cell cycle-related proteins. We also found that the overexpression of AX2R increased the expression of KLF2, mediating VEGF and VEGFR2. Conclusions: Overexpression of AX2R contributes to the inhibition of NV via suppressing KLF2 ubiquitin-dependent protein degradation, which might therefore be a therapeutic option for NV. It could be considered more broadly as an anti-angiogenic agent in the treatment of neovascular-related diseases in the future.

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Section: Discussionsupporting

confidence: 54%

Overexpression of Annexin A2 Receptor Inhibits Neovascularization via the Promotion of Krüppel-Like Transcription Factor 2

Guo

Song

Zhao

et al. 2018

Cell Physiol Biochem

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“…Steady-state levels can be reduced by affecting any level of gene expression including inhibiting its transcription or translation, or by increasing its rate of degradation. Serving as clear examples of the latter, overexpression of the mammalian ubiquitin E3 ligase MKRN1 results in the degradation of the hTERT telomerase subunit (Kim et al 2005) and overexpression of the SMURF2 ubiquitin E3 destabilizes the KLF5 DNA-binding protein (Du et al 2011). Consistent with its identification as a specific regulator, knockdown of the SMURF2 E3 ligase increases the level of KLF5.…”

Section: Inhibitionmentioning

confidence: 51%

Gene Overexpression: Uses, Mechanisms, and Interpretation

2012

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The classical genetic approach for exploring biological pathways typically begins by identifying mutations that cause a phenotype of interest. Overexpression or misexpression of a wild-type gene product, however, can also cause mutant phenotypes, providing geneticists with an alternative yet powerful tool to identify pathway components that might remain undetected using traditional loss-of-function analysis. This review describes the history of overexpression, the mechanisms that are responsible for overexpression phenotypes, tests that begin to distinguish between those mechanisms, the varied ways in which overexpression is used, the methods and reagents available in several organisms, and the relevance of overexpression to human disease.

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“…KLF5 has been identified as an unstable protein that is ubiquitinated by WWP1 [17], FBW7 [18], SMURF2 [15] and EFP [16]. KLF5 K369Q could be detached from the proposed E3 ligase constitutively (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…Our previous studies have shown that KLF5 is an unstable protein with a short half-life [14] and is degraded at least through the ubiquitination-proteasome pathway [14]. WWP1, FBW7, SMURF2 and EFP have been identified as E3 ligases that target KLF5 for ubiquitination and subsequent degradation [15–18]. However, whether the acetylation of KLF5 affects its protein stability is unknown at present.…”

Section: Introductionmentioning

confidence: 99%

HDAC-mediated deacetylation of KLF5 associates with its proteasomal degradation

Tao

Zhang

et al. 2018

Biochemical and Biophysical Research Communications

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Krüppel-like factor 5 (KLF5) is a basic transcription factor that regulates diverse cellular processes during tumor development. Acetylation of KLF5 at lysine 369 (K369) reverses its function from promoting to suppressing cell proliferation and tumor growth. In this study, we examined the regulation of KLF5 by histone deacetylases in the prostate cancer cell line DU 145. While confirming the functions of HDAC1/2 in KLF5 deacetylation and the promotion of cell proliferation, we found that the knockdown of HDAC1/2 upregulated KLF5 protein but not KLF5 mRNA, and the increase in KLF5 protein level by silencing HDAC1/2 was at least in part due to decreased proteasomal degradation. Deacetylase activity was required for HDAC1/2-mediated KLF5 degradation, and mutation of KLF5 to an acetylation-mimicking form prevented its degradation, even though the mutation did not affect the binding of KLF5 with HDAC1/2. Mutation of K369 to arginine, which prevents acetylation, did not affect the binding of KLF5 to HDAC1 or the response of KLF5 to HDAC1/2-promoted degradation. These findings provide a novel mechanistic association between the acetylation status of KLF5 and its protein stability. They also suggest that maintaining KLF5 in a deacetylated form may be an important mechanism by which KLF5 and HDACs promote cell proliferation and tumor growth.

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The E3 Ubiquitin Ligase SMAD Ubiquitination Regulatory Factor 2 Negatively Regulates Krüppel-like Factor 5 Protein

Cited by 44 publications

References 32 publications

Overexpression of Annexin A2 Receptor Inhibits Neovascularization via the Promotion of Krüppel-Like Transcription Factor 2

Overexpression of Annexin A2 Receptor Inhibits Neovascularization via the Promotion of Krüppel-Like Transcription Factor 2

Gene Overexpression: Uses, Mechanisms, and Interpretation

HDAC-mediated deacetylation of KLF5 associates with its proteasomal degradation

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