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Translational Autoregulation of the S. cerevisiae High-affinity Polyamine Transporter Hol1

ABSTRACT: Translational Autoregulation of the S. cerevisiae High-affinity Polyamine Transporter Hol1

PROVIDER: PRJNA719381 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR14135412.fastq.gz	Fastqsanger.gz
	SRR14135413.fastq.gz	Fastqsanger.gz
	SRR14135414.fastq.gz	Fastqsanger.gz
	SRR14135415.fastq.gz	Fastqsanger.gz
	SRR14135416.fastq.gz	Fastqsanger.gz

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Translational Autoregulation of the S. cerevisiae High-affinity Polyamine Transporter Hol1

Project description:Study is aimed to understand translation regulation of mRNA in low and high polyamines in S. cerevisiae. mRNA seq and Ribosome profiling were performed in yeast cells, in duplicates, under low (0 spermidine) and high (10 µM) spermidine. Translation efficiency was calculated (low vs high SPD) and GO analysis reveals 23 transporters as the only class undergo TE downregulation in high SPD condition. Further, we investigated role of these 23 transporters in polyamine transport and identified HOL1 as the high affinity polyamine transporter. This study further show that HOL1 is under polyamine control of translation regulation through conserved fungal uORF MLLLPS*.

2021-08-09 | GSE171392 | GEO

Translational autoregulation of the S. cerevisiae high-affinity polyamine transporter Hol1.

Project description:Polyamines, small organic polycations, are essential for cell viability, and their physiological levels are homeostatically maintained by post-transcriptional regulation of key biosynthetic enzymes. In addition to de novo synthesis, cells can also take up polyamines; however, identifying cellular polyamine transporters has been challenging. Here we show that the S. cerevisiae HOL1 mRNA is under translational control by polyamines, and we reveal that the encoded membrane transporter Hol1 is a high-affinity polyamine transporter and is required for yeast growth under limiting polyamine conditions. Moreover, we show that polyamine inhibition of the translation factor eIF5A impairs translation termination at a Pro-Ser-stop motif in a conserved upstream open reading frame on the HOL1 mRNA to repress Hol1 synthesis under conditions of elevated polyamines. Our findings reveal that polyamine transport, like polyamine biosynthesis, is under translational autoregulation by polyamines in yeast, highlighting the extensive control cells impose on polyamine levels.

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HeLa cell protein interactome of polyamine affinity-based probes.

Project description:HeLa cells were incubated with affinity-based probes designed as synthetic analogs of endogenous metabolites: putrescine, spermidine, and spermine, while a monoamine compound served as a control. Probe-protein interactions were captured in situ using UV-induced crosslinking. After cell lysis, biorthogonal ligation and affinity purification of the probe-conjugated proteins were carried out. These proteins were digested with trypsin, and the resulting peptides were labeled with TMT. Proteins that showed enrichment in samples treated with polyamine probes, compared to those treated with the monoamine control, were identified as polyamine-binding proteins.

2025-09-22 | PXD058706 | Pride

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Project description:Serotonin regulates its own production, degradation and release in the Raphe nuclei of the brain mainly by modifying the level and activity of autoreceptors on serotonin-producing neurons, such as 5-HT1A and 5-HT1B. This autoregulation is probably the reason for the delayed response of patients with depression to the most common antidepressive drugs inhibiting the serotonin transporter (SERT), a molecule responsible for the reuptake of serotonin and therefore for its quick removal from the synaptic cleft. However, the signaling pathways involved in autoregulation and the molecular alterations induced by serotonin on its autoreceptors are yet unclear. We aim at analysing changes in levels of proteins induced by longterm changes of serotonin levels in Raphe nuclei of mice with low and elevated serotonin release in the brain to reveal the signaling pathways involved in the autoregulation of serotonergic neurons.

2025-05-07 | PXD055579 | Pride

Arabidopsis cationic amino acid transporter (CAT) is a high affinity transporter for a systemic herbicide L-Phosphinothricin

Project description:L-phosphinothricin (PPT) is a broad-spectrum herbicide, however its transportation mechanisms remain unidentified. Here we present cationic amino acid transporters (CAT) as the high affinity transporter of PPT, explaining the different sensitivities of flowering and non-flowering plants. Comparative analysis between Arabidopsis thaliana and Marchantia polymorpha revealed non-target site mechanisms for PPT resistance in the latter. LC-MS analysis showed minimal PPT uptake by M. polymorpha, while transcriptomic analysis identified four candidate transporters for PPT uptake. Overexpression of AtCAT1 and AtCAT5 in M. polymorpha increased PPT susceptibility and uptake. Protein molecular dynamics simulation revealed the modes of PPT transportation, allowing for mutagenesis strategies to reduce PPT susceptibility. This study reports the first high affinity PPT transporter, AtCAT, and highlights the natural diversity of PPT susceptibility due to transportation.

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Shotgun analysis of proteins in HeLa cells following treatment with polyamine affinity-based probes

Project description:HeLa cells were incubated with affinity-based probes, synthetic analogs of endogenous metabolites: putrescine, spermidine, and spermine. Cells were photoirradiated briefly and proteins were extracted, digested with trypsin, and the resulting peptides were labeled with TMT. Proteins were quantified across conditions to assess the effect of polyamine analogs on total protein levels in cells. This dataset is related to dataset PXD058706 (HeLa cell protein interactome of polyamine affinity-based probes).

2025-09-22 | PXD062452 | Pride

Arabidopsis cationic amino acid transporter (CAT) is a high affinity transporter for a systemic herbicide L-Phosphinothricin

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Project description:Polyamine uptake transporter 2 is essential for systemic acquired resistance establishment in Arabidopsis.

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Project description:Diffuse intrinsic pontine glioma (DIPG) is an incurable malignant childhood brain tumour, with no active systemic therapies and a 5-year survival of less than 1%. Polyamines are small organic polycations that are essential for DNA replication, translation and cell proliferation. Ornithine decarboxylase 1 (ODC1), the rate limiting enzyme in polyamine synthesis, is irreversibly inhibited by difluoromethylornithine (DFMO). Herein we show that polyamine synthesis is upregulated in DIPG, leading to sensitivity to DFMO. DIPG cells compensate for ODC1 inhibition by upregulation of the polyamine transporter SLC3A2. Treatment with the polyamine transporter inhibitor AMXT 1501 reduced uptake of polyamines in DIPG cells, and co-administration of AMXT 1501 and DFMO led to potent in vitro activity, and significant extension of survival in three aggressive DIPG orthotopic animal models. Collectively, these results demonstrate the potential of dual targeting of polyamine synthesis and uptake as a therapeutic strategy for incurable DIPG.

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Polyamine uptake transporter 2 is essential for systemic acquired resistance establishment in Arabidopsis.

Project description:Plants have evolved a tightly regulated and inducible immune system to resist pathogen attack. After the initial contact with the pathogen, plants are significantly more resistant to future challenges, even from unrelated pathogens, an event known as systemic acquired resistance (SAR). In plants polyamines are involved in the regulation and establishment of the defense response by modulating their metabolism, specifically their biosynthesis, conjugation and catabolism. However, it is not known whether polyamine transport is involved in the plant defense response. Herein, we demonstrate that the PUT2/LAT4 transporter is required to establish local immunity and systemic defense responses in the A. thaliana-Pseudomonas syringae pv. tomato DC3000 (Pst) pathosystem. Our data revealed a dual phenotype in the put2-1 mutant after Pst inoculation, on one side, SAR was abolished in the systemic leaves while on the other, basal resistance to Pst was enhanced. Remarkably, this phenotype persisted even upon inoculation with unrelated pathogens such as Botrytis cinerea, highlighting a broad-spectrum SAR deficiency. To gain deeper insights into the involvement of PA transport in SAR, transcriptomic and metabolomic data of the put2-1 mutant were obtained.

2024-11-04 | GSE275478 | GEO

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