Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Identification of a USP9X Substrate NFX1-123 by SILAC-based Quantitative Proteomics

ABSTRACT: The deubiquitinase USP9X related with multiple diseases including neurodegeneration, epilepsy and various type of tumor, by targeting different substrates. In our study, to deep study USP9X functional candidate substrates, we performed SILAC-based quantitative proteomic to compare knockdown USP9X with wild type HeLa cells to screen USP9X potential substrates. Further, we carried out Flag-NFX1-123 affinity tag-based interaction mass spectrometry method and verified that the X-box binding nuclear factor NFX1-123 is a substrate of USP9X. Further experimental evidences confirmed that USP9X stabilizedNFX1-123 protein level rather than mRNA levels via direct interaction and efficient deubiquitination of NFX1-123 protein

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Keratinocyte

DISEASE(S): Cervix Carcinoma

SUBMITTER: xiangling chen

LAB HEAD: Hu Zhou

PROVIDER: PXD012871 | Pride | 2019-05-13

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	20150422_GJ_U1_1.raw	Raw
	20150422_GJ_U1_10.raw	Raw
	20150422_GJ_U1_11.raw	Raw
	20150422_GJ_U1_12.raw	Raw
	20150422_GJ_U1_13.raw	Raw

Items per page:

1 - 5 of 92

Publications

Identification of a USP9X Substrate NFX1-123 by SILAC-Based Quantitative Proteomics.

Chen Xiangling X Lu Dayun D Gao Jing J Zhu Hongwen H Zhou Yanting Y Gao Daming D Zhou Hu H

Journal of proteome research 20190522 6

The deubiquitinase USP9X is involved in multiple diseases including neurodegeneration, epilepsy, and various types of tumors by targeting different substrates. In the present study, we aimed to explore the potential substrates of USP9X and performed SILAC-based quantitative proteomics to compare these substrates in USP9X-knockdown and wild-type HeLa cells. We consequently carried out Flag-NFX1-123 tag affinity-based mass spectrometry and confirmed that the X-box binding nuclear factor NFX1-123 i ...[more]

PMID: 31059266

Similar Datasets

Loss of Usp9x increases Wnt and Notch signaling in neural progenitors

Project description:Correct neural progenitor fate determination requires the coordination of extrinsic fate determinant signals with intrinsic responses. Post-translational modifications dynamically alter protein function and so are ideally situated to regulate development. Here we show that the deubiquitylaying enzyme, Usp9x modulates both intrinsic and extrinsic regulators of mouse neural progenitors. Nestin-cre mediated deletion of Usp9x from neural progenitors results in a transient disruption of cell adhesion and apical-basal polarity as well as the premature differentiation of intermediate neural progenitors. Ablation of Usp9x also significantly increased Î²-catenin protein levels, especially S33/S37/T41 phospho-Î²-catenin, and Wnt signalling. Usp9x was found to be part of the Î²-catenin destruction complex and loss of Usp9x affects destruction complex composition. Notch signalling was also increased in Usp9x ablated neural progenitors, coinciding with decreased Itch and Numb, and increased Notch intracellular domain protein levels. Usp9x co-localized and immunopreciptiated with Numb from neural progenitors suggesting it is required for Numb stabilisation. These data suggest Usp9x plays a role in coordinating intrinsic responses to extrinsic signals during neural development.

2017-08-18 | E-MTAB-4150 | biostudies-arrayexpress

Ubiquitin remnant analysis in melanoma post inhibition of Usp9x

Project description:Assay the ubiquitinated protein remmants post depletion or inhibition of the deubiquitinase Usp9x in melanoma cells.

2017-02-20 | PXD005417 | Pride

A novel USP9X substrate TTK contributes to tumorigenesis in non-small cell lung cancer

Project description:The X-linked deubiquitinase, USP9X, is implicated in multiple cancers by targeting various substrates. Increased expression of USP9X is observed in non-small cell lung cancer (NSCLC) and is correlated with poor prognosis. However, the molecular mechanism for USP9X regulating tumor cell survival and tumorigenesis in NSCLC is less defined. In this study, chemical labeling quantitative proteomic screening was applied to analyze A549 cells with or without USP9X RNA interference, and the resulting data suggested that TTK is a potential substrate of USP9X. Further experimental evidences confirmed that USP9X stabilized TTK via direct interaction and deubiquitination of TTK. Moreover, knockdown of USP9X or TTK inhibited cell proliferation, migration and tumorigenesis, and the immunohistochemical analysis of clinical NSCLC samples showed that the protein expression levels of USP9X and TTK were significantly elevated and positively correlated in tumor tissues. In summary, our data demonstrated that the USP9X-TTK axis may play a critical role in NSCLC, and could be considered as the potential therapeutic target.

2018-05-07 | PXD007726 | Pride

Identification of mitotic substrates of phospho-regulated USP9X

Project description:We previously identified the deubiquitylase USP9X as a G2/M-specific CDC14B interactor (PXD012732) and subsequently identified USP9X as a specific CDC14B target (PXD012733). To investigate relevant mitotic substrates of phospho-regulated USP9X, we next performed a Stable isotope labeling by amino acids in cell culture (SILAC)-based screen in which ubiquitylated proteins were purified from control or USP9X depleted HEK 293T cells that were either asynchronous or synchronized in mitosis. Stable expression of recombinant His-tagged ubiquitin has been used for selective enrichment of ubiquitinylated proteins in both conditions.

2020-03-25 | PXD012734 | Pride

USP9X phosphopeptide mapping - We previously identified the deubiquitylase USP9X as a G2/M-specific CDC14B interactor (PXD012732). The functional connection of these proteins has been further investigated by a quantitative comparison of USP9X phosphorylation in WT vs CDC14B-overexpressing HEK 293Tcells (ATCC® CRL-3216™). The approach identified USP9X serine 2563 as a specific CDC14B target. Phosphorylation at USP9XS2563 has been confirmed by in-house phosphospecific antibodies.

Project description:We previously identified the deubiquitylase USP9X as a G2/M-specific CDC14B interactor (PXD012732). The functional connection of these proteins has been further investigated by a quantitative comparison of USP9X phosphorylation in WT vs CDC14B-overexpressing HEK 293Tcells (ATCC® CRL-3216™). The approach identified USP9X serine 2563 as a specific CDC14B target. Phosphorylation at USP9XS2563 has been confirmed by in-house phosphospecific antibodies.

2020-03-25 | PXD012733 | Pride

USP9X controls the resolution of stalled ribosomes

Project description:When a ribosome stalls during translation, it runs the risk of collision with a trailing ribosome. Such an encounter leads to the formation of a stable di-ribosome complex, which needs to be resolved by a dedicated machinery. The initial stalling and the subsequent resolution of di-ribosomal complexes requires activity of Makorin and ZNF598 ubiquitin E3 ligases respectively, through ubiquitylation of the eS10 and uS10 sub-units of the ribosome. It is common for the stability of RING E3 ligases to be regulated by an interacting deubiquitylase, which often opposes auto-ubiquitylation of the E3. Here, we show that the deubiquitylase USP9X directly interacts with ZNF598 and regulates its abundance through the control of protein stability. We have developed a highly specific small molecule inhibitor of USP9X. In the absence of USP9X or following chemical inhibition of its catalytic activity, steady state levels of Makorins and ZNF598 are diminished and the ribosomal quality control pathway is impaired.

2021-01-26 | PXD018662 | Pride

NUDT21 Negatively Regulates PSMB2 and CXXC5 by Alternative Polyadenylation and Contributes to Hepatocellular Carcinoma Suppression

Project description:HCC is the third leading cause of cancer-related deaths worldwide. However, the molecular mechanisms underlying the progression of HCC is still largely elusive. NUDT21 (CFIm25) is an important mediator of 3′ UTR APA and demonstrates a causal relationship between alternative polyadenylation and cancer cell proliferation. Although the function of NUDT21 has been explored in glioblastoma tumor, the functional significance of NUDT21 in solid tumors is not well understood. In this study, we observed that NUDT21 is suppressed in human HCC tissues, compared to adjacent noncancerous tissues. In addition we found that the HCC patients with suppressed NUDT21 are statistically associated with poor outcomes. These observations suggest NUDT21 possibly functions as tumor suppressor in hepatocellular carcinoma (HCC).

2018-01-25 | E-MTAB-6368 | biostudies-arrayexpress

The deubiquitinase Usp9x regulates PRC2-mediated chromatin reprogramming during mouse development

Project description:Pluripotent cells of the mammalian embryo undergo extensive chromatin rewiring to prepare for lineage commitment after implantation. Repressive H3K27me3, deposited by Polycomb Repressive Complex 2 (PRC2), is reallocated from large blankets in pre-implantation embryos to mark promoters of developmental genes. The regulation of the global redistribution of H3K27me3 is poorly understood. Here we report a post-translational mechanism that destabilizes PRC2 to constrict H3K27me3 during lineage commitment. Using an auxin-inducible degron system, we show that the deubiquitinase Usp9x is required for mouse embryonic stem (ES) cell self-renewal. Usp9x-high ES cells have high PRC2 levels and bear a chromatin and transcriptional signature of the pre-implantation embryo, whereas Usp9x-low ES cells resemble the post-implantation, gastrulating epiblast. We show that Usp9x interacts with, deubiquitinates and stabilizes PRC2. Deletion of Usp9x in post-implantation embryos results in the derepression of genes that normally gain H3K27me3 after gastrulation, followed by the appearance of morphological abnormalities at E9.5, pointing to a recurrent link between Usp9x and PRC2 during development. Usp9x is a marker of “stemness” and is mutated in various neurological disorders and cancers. Our results unveil a Usp9x-PRC2 regulatory axis that is critical at peri-implantation and may be redeployed in other stem cell fate transitions and disease states.

2021-01-25 | GSE146800 | GEO

USP9X stabilizes EWS-FLI1 to promote cancer cell growth in Ewing’s sarcoma

Project description:Ewing’s sarcoma (EWS) is a cancer of the bones or soft tissues in children and adolescents. EWS-FLI1 is a transcriptional factor and the key driver of EWS. To characterize the changes of downstream transcriptional profiles of EWS-FLI1 in A673 cells upon knockdown of EWS-FLI1 and deubiquitinase USP9X, RNA-seq was performed in A673 cells transfected with EWS-FLI1 esiRNA, USP9X esiRNA or GFP esiRNA as control. The data indicates that USP9X regulates transcriptional profiles of EWS through mediating EWS-FLI1.

2022-01-01 | GSE185429 | GEO

A proteomic approach for systematic mapping of human deubiquitinating enzymes substrates

Project description:In the present study we implemented the bioUb approach to identify DUB substrates in a systemic manner. DUBs have been individually silenced so that the ubiquitination levels of their respective substrates are increased, and hence, upon isolation, susbtrates have been detected by mass spectrometry. Here we report quantitative proteomic data of the putative substrates of 5 human DUBs: USP1, USP7, USP9X, USP11 and USP42.

2021-04-29 | PXD025443 | Pride

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data