The ubiquitin-like modifier FAT10 inhibits retinal PDE6 activity and mediates its proteasomal degradation

Boehm, Annika N; Bialas, Johanna; Catone, Nicola; Sacristán-Reviriego, Almudena; Spuy, Jacqueline van der; Groettrup, Marcus; Aichem, Annette

doi:10.1074/jbc.ra120.013873

Cited by 7 publications

(10 citation statements)

References 55 publications

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“…Of note, recombinant UBA6 alone was sufficient to initiate the conjugate formation between FAT10 and HUWE1 ( Fig 1D , IB: HA, lanes 7 and 19). The addition of USE1 did not further increase the amount of FAT10ylated HUWE1 ( Fig 1D , IB: HA, lanes 9 and 21), as observed already previously for other FAT10 substrates [ 55 – 58 ]. A transfer of ubiquitin onto HUWE1 was not detectable under these conditions ( Fig 1D , IB: HA, lanes 12 and 24).…”

Section: Resultssupporting

confidence: 88%

The ubiquitin-like modifier FAT10 covalently modifies HUWE1 and strengthens the interaction of AMBRA1 and HUWE1

Mueller,

Bialas,

Ryu

et al. 2023

PLoS ONE

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The ubiquitin-like modifier FAT10 is highly upregulated under inflammatory conditions and targets its conjugation substrates to the degradation by the 26S proteasome. This process termed FAT10ylation is mediated by an enzymatic cascade and includes the E1 activating enzyme ubiquitin-like modifier activating enzyme 6 (UBA6), the E2 conjugating enzyme UBA6-specific E2 enzyme 1 (USE1) and E3 ligases, such as Parkin. In this study, the function of the HECT-type ubiquitin E3 ligase HUWE1 was investigated as a putative E3 ligase and/or conjugation substrate of FAT10. Our data provide strong evidence that HUWE1 is FAT10ylated in a UBA6 and FAT10 diglycine-dependent manner in vitro and in cellulo and that the HUWE1-FAT10 conjugate is targeted to proteasomal degradation. Since the mutation of all relevant cysteine residues within the HUWE1 HECT domain did not abolish FAT10 conjugation, a role of HUWE1 as E3 ligase for FAT10ylation is rather unlikely. Moreover, we have identified the autophagy-related protein AMBRA1 as a new FAT10 interaction partner. We show that the HUWE1-FAT10 conjugate formation is diminished in presence of AMBRA1, while the interaction between AMBRA1 and HUWE1 is strengthened in presence of FAT10. This implies a putative interplay of all three proteins in cellular processes such as mitophagy.

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Section: Resultssupporting

confidence: 88%

The ubiquitin-like modifier FAT10 covalently modifies HUWE1 and strengthens the interaction of AMBRA1 and HUWE1

Mueller,

Bialas,

Ryu

et al. 2023

PLoS ONE

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“…The effects of PDE6H depletion could be further supported by overexpression of the gene in knockout cells. Epitope tagging of the inhibitory PDE6 could also help but to ensure specific tagging of PDE6γ′ only, and not PDE6γ; the N-terminal of PDE6γ′ needs to be targeted, which has not previously been done [ 81 , 82 ]. As the crystal structure of the N-terminal of PDE6γ′, despite its known contribution to PDE6 complex formation and activity [ 83 ], has not been fully determined, this would be difficult to interpret.…”

Section: Discussionmentioning

confidence: 99%

Cone photoreceptor phosphodiesterase PDE6H inhibition regulates cancer cell growth and metabolism, replicating the dark retina response

Yalaz,

Bridges,

Alham

et al. 2024

Cancer Metab

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Background PDE6H encodes PDE6γ′, the inhibitory subunit of the cGMP-specific phosphodiesterase 6 in cone photoreceptors. Inhibition of PDE6, which has been widely studied for its role in light transduction, increases cGMP levels. The purpose of this study is to characterise the role of PDE6H in cancer cell growth. Methods From an siRNA screen for 487 genes involved in metabolism, PDE6H was identified as a controller of cell cycle progression in HCT116 cells. Role of PDE6H in cancer cell growth and metabolism was studied through the effects of its depletion on levels of cell cycle controllers, mTOR effectors, metabolite levels, and metabolic energy assays. Effect of PDE6H deletion on tumour growth was also studied in a xenograft model. Results PDE6H knockout resulted in an increase of intracellular cGMP levels, as well as changes to the levels of nucleotides and key energy metabolism intermediates. PDE6H knockdown induced G1 cell cycle arrest and cell death and reduced mTORC1 signalling in cancer cell lines. Both knockdown and knockout of PDE6H resulted in the suppression of mitochondrial function. HCT116 xenografts revealed that PDE6H deletion, as well as treatment with the PDE5/6 inhibitor sildenafil, slowed down tumour growth and improved survival, while sildenafil treatment did not have an additive effect on slowing the growth of PDE6γ′-deficient tumours. Conclusions Our results indicate that the changes in cGMP and purine pools, as well as mitochondrial function which is observed upon PDE6γ′ depletion, are independent of the PKG pathway. We show that in HCT116, PDE6H deletion replicates many effects of the dark retina response and identify PDE6H as a new target in preventing cancer cell proliferation and tumour growth.

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“…The role of ubiquitination in ophthalmology has been studied in several ways. In a study by Fu SH et al [ 18 ], the epithelial-mesenchymal transition and cell permeability of retinal pigment epithelial cells were discovered to be impacted by the ubiquitination degradation process, which has an impact on diabetic retinopathy. In a study by Annika N Boehm et al [ 19 ], it was discovered that in inflammatory eye diseases, the human leukocyte antigen (HLA)-F adjacent transcript 10 (FAT10) family of ubiquitin-like modifiers can lead to the loss of phosphodiesterase 6 (PDE6) by targeting PDE6 for proteasomal degradation through the formation of covalent covalent bonds.…”

Section: Discussionmentioning

confidence: 99%

Identification of potential biomarkers of myopia based on machine learning algorithms

Zhang,

Wang,

Wang

et al. 2023

BMC Ophthalmol

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Purpose This study aims to identify potential myopia biomarkers using machine learning algorithms, enhancing myopia diagnosis and prognosis prediction. Methods GSE112155 and GSE15163 datasets from the GEO database were analyzed. We used “limma” for differential expression analysis and “GO plot” and “clusterProfiler” for functional and pathway enrichment analyses. The LASSO and SVM-RFE algorithms were employed to screen myopia-related biomarkers, followed by ROC curve analysis for diagnostic performance evaluation. Single-gene GSEA enrichment analysis was executed using GSEA 4.1.0. Results The functional analysis of differentially expressed genes indicated their role in carbohydrate generation and polysaccharide synthesis. We identified 23 differentially expressed genes associated with myopia, four of which were highly effective diagnostic biomarkers. Single gene GSEA results showed these genes control the ubiquitin-mediated protein hydrolysis pathway. Conclusion Our study identifies four key myopia biomarkers, providing a foundation for future clinical and experimental validation studies.

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The ubiquitin-like modifier FAT10 inhibits retinal PDE6 activity and mediates its proteasomal degradation

Cited by 7 publications

References 55 publications

The ubiquitin-like modifier FAT10 covalently modifies HUWE1 and strengthens the interaction of AMBRA1 and HUWE1

The ubiquitin-like modifier FAT10 covalently modifies HUWE1 and strengthens the interaction of AMBRA1 and HUWE1

Cone photoreceptor phosphodiesterase PDE6H inhibition regulates cancer cell growth and metabolism, replicating the dark retina response

Identification of potential biomarkers of myopia based on machine learning algorithms

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