Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Total proteome analysis of NUDT5 PROTACs

ABSTRACT: 6-Thioguanine (6-TG), an FDA-approved antimetabolite drug, is widely used in the treatment of leukemia. Its cellular effects require metabolic activation and are regulated through interactions with various proteins such as NUDT15, which catalyzes the hydrolysis of the active 6-TG metabolites 6-thio-deoxyGTP (6-thio-dGTP) and 6-thio-GTP. Recent genome-wide CRISPR loss-of-function studies have identified another NUDIX hydrolase, NUDT5, as a crucial mediator of 6-TG toxicity. Here, we present the development and characterization of potent and selective NUDT5 degraders, guided by a cell-based assay screening strategy. These degraders, in conjunction with orthogonal CRISPR knock-out and reconstitution experiments, reveal a novel and unexpected, non-enzymatic role for NUDT5 in modulating the cellular response to 6-TG. Depletion of NUDT5 protein is antagonistic to NUDT15 inhibition, suggesting a distinct mode-of-action with potential implications for patient therapy.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Ludwig Bauer

LAB HEAD: Kilian Huber

PROVIDER: PXD068003 | Pride | 2026-05-13

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	EC_147-300ng-31_DMSO_1.raw	Raw
	EC_147-300ng-32_DMSO_2.raw	Raw
	EC_147-300ng-33_DMSO_3.raw	Raw
	EC_147-300ng-34_DMSO_4.raw	Raw
	EC_147-300ng-35_ASM127_1.raw	Raw

Items per page:

1 - 5 of 30

Similar Datasets

Large-scale interaction proteomic profiling of NUDT5 and PPAT interactors

Project description:Folate metabolism is intricately linked to purine de novo synthesis through the incorporation of folate-derived one-carbon units into the purine scaffold. Here, we investigate the chemical and genetic dependencies caused by mutations in the folate enzyme MTHFD1 and discover a key role for Nudix hydrolase 5 (NUDT5) in regulating purine de novo synthesis. Through genetic knockout and development of a selective chemical NUDT5 degrader, we uncover an unprecedented scaffolding role rather than NUDT5 enzymatic activity is responsible for this phenotype. We find that NUDT5 interacts with the rate-limiting enzyme of purine de novo synthesis, PPAT, to repress the pathway in response to elevated purine levels. Our findings establish NUDT5 as an important regulator of purine de novo synthesis and elucidate its role in mediating sensitivities to purine analogs in cancer treatment and to adenosine in MTHFD1 deficiency.

2025-11-03 | PXD067706 | Pride

NUDT5 regulates purine metabolism and thiopurine sensitivity by interacting with PPAT

Project description:WT-Mock-1: WT cells treated with vehicle for 24 hours, replicate-1: 126 WT-Mock-2: WT cells treated with vehicle for 24 hours, replicate-2: 127N WT-Mock-3: WT cells treated with vehicle for 24 hours, replicate-3: 127C WT-Mock-4: WT cells treated with vehicle for 24 hours, replicate-4: 128N WT-6-TG-1: WT cells treated with 0.5ug/mL 6-TG for 24 hours , replicate-1: 128C WT-6-TG-2: WT cells treated with 0.5ug/mL 6-TG for 24 hours , replicate-2: 129N WT-6-TG-3: WT cells treated with 0.5ug/mL 6-TG for 24 hours , replicate-3: 129C WT-6-TG-4: WT cells treated with 0.5ug/mL 6-TG for 24 hours , replicate-4: 130N NUDT5-Mock-1: NUDT5 KO cells treated with vehicle for 24 hours, replicate-1: 130C NUDT5-Mock-2: NUDT5 KO cells treated with vehicle for 24 hours, replicate-2: 131N NUDT5-Mock-3: NUDT5 KO cells treated with vehicle for 24 hours, replicate-3: 131C NUDT5-Mock-4: NUDT5 KO cells treated with vehicle for 24 hours, replicate-4: 132N NUDT5-6-TG-1: NUDT5 KO cells treated with 0.5ug/mL 6-TG for 24 hours , replicate-1: 132C NUDT5-6-TG-2: NUDT5 KO cells treated with 0.5ug/mL 6-TG for 24 hours , replicate-2: 133N NUDT5-6-TG-3: NUDT5 KO cells treated with 0.5ug/mL 6-TG for 24 hours , replicate-3: 133C NUDT5-6-TG-4: NUDT5 KO cells treated with 0.5ug/mL 6-TG for 24 hours , replicate-4: 134N

2025-03-24 | MSV000097432 | MassIVE

Genome wide analsis of the chromatin accessibility and gene expression in control and NUDT5 depleted U2OS cells

Project description:NUDT5 was reported to generate ATP to support local chromatin remodeling and transcription of hormone-responsive genes. Our research has found that NUDT5 is also required for DNA damage repair by homologous recombination. To investigate how NUDT5 regulates HR, we depleted NUDT5 in U2OS cells by siRNA and compared the transcriptome profiling (RNA-seq) of NUDT5 depleted cells with control cells to investigate whether NUDT5 regulate HR by altering the expression of essential components of the HR machinery. The results suggest that NUDT5 does not regulate HR by altering the expression of essential components of the HR machinery. To investigate whether NUDT5 contributes to chromatin remodeling after DNA damage, we compared the chromatin accessibility profiling (ATAC-seq) in NUDT5 depleted U2OS cells and control cells, either before or after ionizing radiation. The results suggest that NUDT5 silencing blocked most DNA damage-induced changes in chromatin accessibility following radiation exposure and implies that NUDT5 may support the activity of remodeling ATPases during damage repair.

2023-02-14 | GSE196718 | GEO

NUDT5 restrains PPAT to promote pyrimidine synthesis and effective nucleobase therapy - NUDT5 co-immunoprecipitation

Project description:Nucleotides are essential building blocks for nucleic acid synthesis, signaling, and metabolism. Rapidly proliferating cells require large amounts of nucleotides, making nucleotide metabolism a widely exploited target for cancer therapy. However, resistance frequently emerges, highlighting the need for a deeper understanding of nucleotide regulation. Here, we use uridine-sensitized CRISPR-Cas9 screening to reveal novel regulators of pyrimidine synthesis. We identify NUDT5 which we show binds to and negatively regulates PPAT, the rate-limiting enzyme in purine biosynthesis. We demonstrate the NUDT5-PPAT interaction prevents excessive purine production, maintains phosphoribosyl pyrophosphate (PRPP), a critical substrate for pyrimidine synthesis, and thus enhances conversion of nucleobase analogs into active anti-cancer molecules. We further report that NUDT5 regulates PPAT independently of its enzymatic activity and can be displaced by PRPP, revealing intricate allosteric regulation. Our findings reveal a previously unrecognized mechanism for maintaining nucleotide balance and positions NUDT5 as a potential therapeutic target for overcoming resistance to chemotherapy in proliferative diseases.

2025-10-29 | PXD060353 | Pride

Genome wide analsis of the chromatin accessibility and gene expression in control and NUDT5 depleted U2OS cells [RNA-seq]

2023-02-14 | GSE196722 | GEO

Genome wide analsis of the chromatin accessibility and gene expression in control and NUDT5 depleted U2OS cells [ATAC-seq]

2023-02-14 | GSE196721 | GEO

NUDT5 restrains PPAT to promote pyrimidine synthesis and effective nucleobase therapy - sgNUDT5 proteomics

2025-10-29 | PXD060320 | Pride

BFP pulldown of BFP-NUDT5 HAP1 MTHFD1-K56R cells

Project description:We performed BFP pulldowns with BFP-tagged NUDT5 in HAP1 harboring the MTHFD1 K56R mutation to find interactors of NUDT5. We performed the study under various medium conditions, including full IMDM medium (FULL), IMDM with dialysed FBS (DIA) and DIA supplemented with 50µM adenosine to investigate potential changes of interactors upon different medium conditions. Additionally, we performed Pulldowns with a catalytic inactive NUDT5 variant (E112Q), to check whether its catalytic activity has an influence in the interactome. As a control, we used HAP1 MTHFD1 K56R NUDT5 KO cells. Experiments were performed in duplicates

2025-11-07 | PXD067739 | Pride

MITF deficiency accelerates GNAQ-driven uveal melanoma

Project description:Cutaneous melanoma (CM) and uveal melanoma (UM) both originate from the melanocytic lineage but are primarily driven by distinct oncogenic drivers, BRAF/NRAS or GNAQ/GNA11 respectively. The melanocytic master transcriptional regulator, MITF, is essential for both CM development and maintenance, but its role in UM is largely unexplored. Here, we use zebrafish models to dissect the key UM oncogenic signaling events, and establish the role of MITF in UM tumors. Remarkably, mitfa deficiency was profoundly UM promoting, dramatically accelerating the onset and progression of tumors induced by Tg(mitfa:GNAQQ209L);tp53M214K/M214K. To further explore the role of MITF in GNAQ-driven tumorigenesis, we performed phospho-proteomics and total proteomics on 5 zebrafish GNAQ Tg(mitfa:GNAQQ209L);tp53M214K/M214K tumors and 5 zebrafish GNAQ Tg(mitfa:GNAQQ209L);tp53M214K/M214K;mitfa-/- tumors.

2023-03-11 | PXD030527 | Pride

SMARCA2/4 degradation tails mediate recruitment of two different E3 ligases 

Project description:In this work, we characterise novel SMARCA2/4 degraders using a robust cell biology and structural workflow and reveal for the first time, the recruitment of two E3 ligase substrate receptors: FBXO22 and DCAF16 by the same degradation tail. Furthermore, we were able to chemically fine-tune this dual ligase dependency dialling out DCAF16 providing insights into the mechanisms for DCAF16 and FBXO22 covalent recruitment. Dual ligase recruitment may serve to overcome E3 ligase acquired resistance that may feature with clinical use of degraders.

2026-05-16 | PXD065445 | 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