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Identification of the altered protein between wildtype and YTHDF1 knockout B16/F10 using Tandem Mass Tag

ABSTRACT: The recently described role of RNA methylation in regulating immune cell infiltration into tumors has attracted interest, given its potential impact on immunotherapy response. YTHDF1 is a versatile and powerful m6A reader, but the understanding of its impact on immune evasion is limited. Here, we performed proteomics to reveal the altered proteins between wildtype and YTHDF1 knockout group.

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Melanocyte

DISEASE(S): Melanoma

SUBMITTER: Wanzun Lin

LAB HEAD: Wanzun Lin

PROVIDER: PXD036931 | Pride | 2022-12-06

REPOSITORIES: Pride

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			Action	DRS
	QE_HFX_20220724_50cm_90min_TMT10Plex_F1.raw	Raw
	QE_HFX_20220724_50cm_90min_TMT10Plex_F10.raw	Raw
	QE_HFX_20220724_50cm_90min_TMT10Plex_F11.raw	Raw
	QE_HFX_20220724_50cm_90min_TMT10Plex_F12.raw	Raw
	QE_HFX_20220724_50cm_90min_TMT10Plex_F2.raw	Raw

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Publications

Identification of molecular subtypes based on inflammatory response in lower-grade glioma.

Lin Wanzun W Gao Jing J Zhang Haojiong H Chen Li L Qiu Xianxin X Huang Qingting Q Hu Jiyi J Kong Lin L Lu Jiade J JJ

Inflammation and regeneration 20221001 1

<h4>Background</h4>Inflammatory response is an important characteristic affecting prognosis and therapeutic response in lower-grade glioma (LGG). However, the molecular subtypes based on inflammatory response are still under exploitation.<h4>Methods</h4>The RNA sequencing, somatic mutation, and corresponding clinical data from 1205 LGG patients were obtained from the TCGA, CGGA, and Rembrandt cohorts. Consensus clustering was performed to identify molecular subtypes associated with inflammation. ...[more]

PMID: 36180938

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TMT proteomics analysis of B16/F10

Project description:The recently described role of RNA methylation in regulating immune cell infiltration into tumors has attracted interest, given its potential impact on immunotherapy response. YTHDF1 is a versatile and powerful m6A reader, but the understanding of its impact on immune evasion is limited. Here, we revealed that tumor-intrinsic Ythdf1 drives immune evasion and immune checkpoint inhibitor (ICI) resistance. TMT proteomics analysis was performed to identify the altered protein.

2022-12-06 | PXD036938 | Pride

Proteomic investigation into the action mechanism of inhibitor ptsc1 against Streptococcus pneumoniae

Project description:Phenyl-thiazolylurea-sulfonamides compound 1 (ptsc1) is a new and efficient bacterial inhibitor. Although the inhibitor has antibacterial activity against several model bacteria, the details of its mechanism in Streptococcus pneumoniae (S. pneumoniae) have not been revealed. In the current study, we found that ptsc1 can significantly inhibit the growth of S. pneumoniae. To further investigate the molecular mechanism of its antibacterial effects, quantitative proteomics was performed to identify differential expressed proteins in S. pneumoniae. Compared with the normal group, the expression levels of 38 proteins were upregulated, whereas those of 88 proteins were downregulated (> 1.3 fold-change) in the experimental group (P < 0.05). KEGG pathways analysis revealed that ptsc1 regulated proteins were mainly involved in carbohydrate metabolism, DNA replication and repair, pyrimidine metabolism, fatty acid biosynthesis and oxidative phosphorylation pathways. The results of GO enrichment analysis showed that the inhibitor could lead to dysregulation of bacterial translation and energy metabolism, and cause intracellular oxidative stress.

2022-01-12 | PXD030534 | Pride

Quantitative proteomic analysis of tumour tissues isolated from early and late stage human cervical cancer patients

Project description:Human papillomavirus (HPV) infection related malignancy remains a severe public health problem worldwide, although HPV prophylactic vaccine has been introduced 15 years ago . HPV-associated cancers comprise 29.1% of all 2.2 million infection-related cancers, including nearly 100% of cervical cancers1 and some head and neck cancers . Cervical cancers are the 3rd most common cancer in women worldwide, HPV16 and 18 account for about 70% of cervical cancers. Moreover, high-risk HPV infection, especially HPV16 infection, is related to a proportion of head and neck epithelial carcinoma in both developed and undeveloped countries. HPV related cancers are most severe in developing countries where HPV prophylactic vaccination rates are low.In this project, we use iTRAQ8plex-labelling proteomics to comparatively study the protein contents in the tumour tissues of early and late stage cervical cancer patietns.

2023-01-26 | PXD029103 | Pride

YTHDF1 links hypoxia adaptation and non-small cell lung cancer progression

Project description:10plex TMT labelling quantification expriments were performed to access the knockdown effect of YTHDF1 in cells.3 shGFPs control samples, 3 YTHDF1 shRNA-2 knockdown samples , 3 YTHDF1 shRNA-3 knockdown samples and 1 mixture sample were labelled by TMT 10-plex reagents.

2019-09-27 | PXD015182 | Pride

Identification of specific proteins and metabolic networks associated with sucrose accumulation in sugarcane

Project description:The rate of improvement of sugar content in sugarcane remains low for decades worldwide. Our previous transcriptome studies provided an atlas of sucrose accumulation-related gene expression but little is known about proteins involved. Here this knowledge gap is addressed by a proteomic analysis of experimentally altered sucrose accumulation in sugarcane. Analysis of stem proteomes of ripener ethephon-treated high- and low-sugar genotypes had identified 2983 proteins of which 139 were significantly differentially expressed (DEPs). These DEPs were found to be involved in sugar metabolism-related processes with 25 of them may have a regulatory role in sucrose accumulation. The key proteins identified include UDP-glucose 6-dehydrogenase associated with amino sugar and nucleotide sugar metabolism; those involved in carbon fixation; and fructokinase, β-D-glucosidase and α-glucan phosphorylase involved in starch and sucrose metabolism. Distinct genotype- and ethephon-dependent DEP expression was evident providing new insights on one of the most intractable sugarcane traits to breeding.

2022-08-12 | PXD024569 | Pride

Human intestine LC-MSMS - Transcriptional and proteomic analysis revealed a synergistic effect of aflatoxin M1 and ochratoxin A mycotoxins on intestinal epithelial integrity of differentiated human Caco-2 cells

Project description:Aflatoxin M1 (AFM1) is a common mycotoxin in dairy milk and it is typically concurrently present with other mycotoxins that may represent a threat for food safety. However, knowledge on how AFM1, alone or in combination with other mycotoxins may affect human intestinal epithelial integrity remain to be established. We employed transcriptome and proteome analysis integrated with biological validation to reveal the molecular basis underlining the effect AFM1 and/or ochratoxin A (OTA) exposure on intestinal epithelial integrity of differentiated Caco-2 cells. Exposure to 4 μg/ml of OTA was found to disrupt human gut epithelial integrity, whereas 4 μg/ml of AFM1 did not. Integrated transcriptome and proteome analysis of AFM1 and OTA, alone or in combination, indicate synergistic effect of the two mycotoxins in disrupting intestinal integrity. This effect was mechanistically linked to a broad ranges of pathways related to intestinal integrity enriched by down-regulated genes and proteins, associated to focal adhesion, adherens junction, and gap junction pathways. Furthermore, the cross–omics analysis of mixed AFM1 and OTA compared with OTA alone suggest that kinases family members, including MLCK, MAPKs, and PKC are the potential key regulators on modulating intestinal epithelial integrity. These findings provide novel insight into the synergistic detrimental role of multiple mycotoxins in disrupting intestinal integrity, and, therefore, identify potential target to improve milk safety related to human health.

2018-08-08 | PXD009437 | Pride

Proteomic of huh7 grown in normal and glucose-deficient conditions

Project description:Tumor cell metabolic plasticity is essential for tumor progression and therapeutic response, but the mechanism for regulation of metabolic plasticity remain poorly explored. Here, we identify PROX1 as an essential determinant for tumor metabolic plasticity. Notably, PROX1 is significantly reduced in response to metabolic stress or AMPK activation and is elevated in LKB1-deficient tumors in mice and human. Furthermore, the Ser79 phosphorylation of PROX1 by AMPK significantly alters its protein activity and allows a rapid recruitment of CUL4-DDB1 E3 ubiquitin ligase to promote PROX1 degradation under glucose starvation, a critical event that drives BCAA metabolism rewiring to suppress mTOR signaling. Importantly, PROX1 loss or Ser79 phosphorylation in HCC shows therapeutic resistance to metformin. Consistently, genetic ablation of PROX1 renders LKB1‐deficient KRAS-driven lung cancer resistant to phenformin treatment. Conversely, mice harboring non-phosphorylated PROX1 mutant or LKB1 mutant exhibit high PROX1 stabilization, promoting liver and lung cancer progression. Clinically, AMPK-PROX1 axis in human cancers is important for patient clinical outcomes. Collectively, our results demonstrate that the deficiency in the LKB1-AMPK axis in cancers reactivates PROX1 to sustain intracellular BCAA pools, resulting in enhanced mTOR signaling, and facilitating tumorigenesis and aggressiveness.

2022-09-27 | PXD036977 | Pride

Targeting parvalbumin promotes M2 macrophage polarization energy expenditure in diet-induced mouse obesity models

Project description:Exercise benefits M2 macrophage polarization, energy homeostasis and protects against obesity partially through exercise-induced circulating factors. Here, by unbiased quantitative proteomics on serum samples from sedentary and exercised mice, we identify parvalbumin as a circulating factor suppressed by exercise. Parvalbumin functions as a non-competitive CSF1R antagonist to inhibit M2 macrophage activation and energy expenditure in adipose tissue. More importantly, serum concentrations of parvalbumin positively correlate with obesity in mouse and human, while treating mice with a recombinant parvalbumin blocker prevents its interaction with CSF1R and promotes M2 macrophage polarization and ameliorates diet-induced obesity. Thus, although further studies are required to assess the significance of parvalbumin in mediating the effects of exercise, our results implicate parvalbumin as a potential therapeutic strategy against obesity.

2022-05-10 | PXD033582 | Pride

mRNA-seq of neuronal compartments in shRNA-mediated knockdown of Ythdf1, -2 and -3 in mouse primary cortical neurons (mPCN)

Project description:To study the effect of m6A modifications on subcellular mRNA localization we depleted m6A readers Ythdf1, -2 and -3 with shRNAs from mouse primary cortical neurons (mPCN) and sequenced neuritic and somatic compartments in parallel with scrambled shRNA control.

2023-07-12 | E-MTAB-11569 | biostudies-arrayexpress

N6-methyladenosine Modulates Messenger RNA Translation Efficiency

Project description:N6-methyladenosine (m6A) is the most abundant internal messenger (mRNA) modification in mammalian mRNA. This modification is reversible and non-stoichiometric, which potentially adds an additional layer of variety and dynamic control of mRNA metabolism. The m6A-modified mRNA can be selectively recognized by the YTH family “reader” proteins. The preferential binding of m6A-containing mRNA by YTHDF2 is known to reduce the stability of the target transcripts; however, the exact effects of m6A on translation has yet to be elucidated. Here we show that another m6A reader protein, YTHDF1, promotes ribosome loading of its target transcripts. YTHDF1 forms a complex with translation initiation factors to elevate the translation efficiency of its bound mRNA. In a unified mechanism of translation control through m6A, the YTHDF2-mediated decay controls the lifetime of target transcripts; whereas, the YTHDF1-based translation promotion increases the translation efficiency to ensure effective protein production from relatively short-lived transcripts that are marked by m6A. PAR-CLIP and RIP was used to identify YTHDF1 binding sites followed by ribosome profling and RNA seq to assess the consequences of YTHDF1 siRNA knock-down

2015-06-09 | E-GEOD-63591 | biostudies-arrayexpress

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