PHD3 Regulates p53 Protein Stability by Hydroxylating Proline 359

Rodríguez, Javier; Herrero, Ana; Li, Shuijie; Rauch, Nora; Quintanilla, Andrea; Wynne, Kieran; Krstić, Aleksandar; Acosta, Juan Carlos; Taylor, Cormac T.; Schlisio, Susanne; Kriegsheim, Alex von

doi:10.1016/j.celrep.2018.06.108

Cited by 56 publications

(48 citation statements)

References 51 publications

(62 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is quite common that several deubiquitinating enzymes, working like isozymes, regulate the same substrate under different circumstances, especially in the regulation of vital transcription factors. The stability and ubiquitination of p53 are modulated by diverse deubiquitinating enzymes under different cancer types and stimuli, such as USP42, USP49, PHD3, OTUD1, OTUD5, ATXN3 [62][63][64][65][66][67]. The protein level of Snail is also reported to be precisely regulated by ubiquitination and deubiquitination [32].…”

Section: Discussionmentioning

confidence: 99%

EIF3H promotes aggressiveness of esophageal squamous cell carcinoma by modulating Snail stability

Guo

Zhu

Luo

et al. 2020

J Exp Clin Cancer Res

View full text Add to dashboard Cite

Background Overexpression of eukaryotic translation initiation factor 3H (EIF3H) predicts cancer progression and poor prognosis, but the mechanism underlying EIF3H as an oncogene remains unclear in esophageal squamous cell carcinoma (ESCC). Methods TCGA database and the immunohistochemistry (IHC) staining of ESCC samples were used and determined the upregulation of EIF3H in ESCC. CCK8 assay, colony formation assay and transwell assay were performed to examine the ability of cell proliferation and mobility in KYSE150 and KYSE510 cell lines with EIF3H overexpression or knockdown. Xenograft and tail-vein lung metastatic mouse models of KYSE150 cells with or without EIF3H knockdown were also used to confirm the function of EIF3H on tumor growth and metastasis in vivo. A potential substrate of EIF3H was screened by co-immunoprecipitation assay (co-IP) combined with mass spectrometry in HEK293T cells. Their interaction and co-localization were confirmed using reciprocal co-IP and immunofluorescence staining assay. The function of EIF3H on Snail ubiquitination and stability was demonstrated by the cycloheximide (CHX) pulse-chase assay and ubiquitination assay. The correlation of EIF3H and Snail in clinical ESCC samples was verified by IHC. Results We found that EIF3H is significantly upregulated in esophageal cancer and ectopic expression of EIF3H in ESCC cell lines promotes cell proliferation, colony formation, migration and invasion. Conversely, genetic inhibition of EIF3H represses ESCC tumor growth and metastasis in vitro and in vivo. Moreover, we identified EIF3H as a novel deubiquitinating enzyme of Snail. We demonstrated that EIF3H interacts with and stabilizes Snail through deubiquitination. Therefore, EIF3H could promote Snail-mediated EMT process in ESCC. In clinical ESCC samples, there is also a positive correlation between EIF3H and Snail expression. Conclusions Our study reveals a critical EIF3H-Snail signaling axis in tumor aggressiveness in ESCC and provides EIF3H as a promising biomarker for ESCC treatment.

show abstract

Section: Discussionmentioning

confidence: 99%

EIF3H promotes aggressiveness of esophageal squamous cell carcinoma by modulating Snail stability

Guo

Zhu

Luo

et al. 2020

J Exp Clin Cancer Res

View full text Add to dashboard Cite

show abstract

“…Targeted Quantification of BRD4 Proline Hydroxylation-To measure the change in proline hydroxylation abundance, the Flag-BRD4 vector was expressed in 293T cells and cotransfected with either HA-PHD1, HA-PHD2 or myc-PHD3 vectors or treated with 1% oxygen using a hypoxia chamber (Biospherix Ltd, Parish, NY). To measure the change in proline hydroxylation on PHD2 knockdown, 293T cells stably expressing control shRNA and PHD2 shRNA were generated as previously described (31). Cells were lysed and BRD4 was immunoprecipitated using either anti-flag agarose beads or with 1 g BRD4 antibody (Millipore) and protein A/G beads as previously described (32).…”

Section: Generation Of Stable Knockdown Cell Lines-mentioning

confidence: 99%

Targeted and Interactome Proteomics Revealed the Role of PHD2 in Regulating BRD4 Proline Hydroxylation

Erber¹,

Luo²,

Chen³

2019

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

Targeted quantification assays using MS-based parallel-reaction monitoring and biochemical analysis revealed PHD2 as a key regulatory enzyme of BRD4 proline hydroxylation. Quantitative MS-based interactome and biochemical analyses revealed the functional significance of the proline hydroxylation modification pathway in BRD4-mediated protein-protein interactions and transcriptional activation. Taken together, these results provided mechanistic insights into the oxygen-dependent modification of BRD4 and revealed new roles of the proline hydroxylation pathway in regulating BRD4-mediated protein-protein interaction and gene expression.

show abstract

“…Conflicting data exist relating to whether or not p53 protein expression is induced [96,97,98] or repressed [99,100] by hypoxia. Interestingly, recent work by Rodriguez and colleagues in which they showed the interaction of PHD3 and p53 may shed light on this controversy [51]. Using MS and pharmacological trapping strategies, this study reports that PHD3 hydroxylates p53 at proline 359, which regulates p53 protein stability through the modulation of ubiquitination [51].…”

Section: Evidence For Non-hif Hydroxylation Targetsmentioning

confidence: 99%

“…The tumor suppressor gene, p53, is important in the regulation of a multitude of cellular responses including apoptosis, senescence, DNA repair, and cell cycle arrest [51,80,93,94,95]. Conflicting data exist relating to whether or not p53 protein expression is induced [96,97,98] or repressed [99,100] by hypoxia.…”

Section: Evidence For Non-hif Hydroxylation Targetsmentioning

confidence: 99%

Protein Hydroxylation by Hypoxia-Inducible Factor (HIF) Hydroxylases: Unique or Ubiquitous?

Strowitzki

Cummins

Taylor

2019

Cells

Self Cite

163

146

View full text Add to dashboard Cite

All metazoans that utilize molecular oxygen (O2) for metabolic purposes have the capacity to adapt to hypoxia, the condition that arises when O2 demand exceeds supply. This is mediated through activation of the hypoxia-inducible factor (HIF) pathway. At physiological oxygen levels (normoxia), HIF-prolyl hydroxylases (PHDs) hydroxylate proline residues on HIF-α subunits leading to their destabilization by promoting ubiquitination by the von-Hippel Lindau (VHL) ubiquitin ligase and subsequent proteasomal degradation. HIF-α transactivation is also repressed in an O2-dependent way due to asparaginyl hydroxylation by the factor-inhibiting HIF (FIH). In hypoxia, the O2-dependent hydroxylation of HIF-α subunits by PHDs and FIH is reduced, resulting in HIF-α accumulation, dimerization with HIF-β and migration into the nucleus to induce an adaptive transcriptional response. Although HIFs are the canonical substrates for PHD- and FIH-mediated protein hydroxylation, increasing evidence indicates that these hydroxylases may also have alternative targets. In addition to PHD-conferred alterations in protein stability, there is now evidence that hydroxylation can affect protein activity and protein/protein interactions for alternative substrates. PHDs can be pharmacologically inhibited by a new class of drugs termed prolyl hydroxylase inhibitors which have recently been approved for the treatment of anemia associated with chronic kidney disease. The identification of alternative targets of HIF hydroxylases is important in order to fully elucidate the pharmacology of hydroxylase inhibitors (PHI). Despite significant technical advances, screening, detection and verification of alternative functional targets for PHDs and FIH remain challenging. In this review, we discuss recently proposed non-HIF targets for PHDs and FIH and provide an overview of the techniques used to identify these.

show abstract

PHD3 Regulates p53 Protein Stability by Hydroxylating Proline 359

Cited by 56 publications

References 51 publications

EIF3H promotes aggressiveness of esophageal squamous cell carcinoma by modulating Snail stability

EIF3H promotes aggressiveness of esophageal squamous cell carcinoma by modulating Snail stability

Targeted and Interactome Proteomics Revealed the Role of PHD2 in Regulating BRD4 Proline Hydroxylation

Protein Hydroxylation by Hypoxia-Inducible Factor (HIF) Hydroxylases: Unique or Ubiquitous?

Contact Info

Product

Resources

About