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Regulated biogenesis and targeting of monotopic squalene monooxygenase requires the GET pathway

ABSTRACT: This study investigated get3 mutant yeast cell. Proteins of the GET pathway are involved in targeting of C-terminally anchored transmembrane proteins and protection against lipotoxicity. Get3 cells revealed an altered ergosterol production and susceptibility towards a sterol synthesis inhibiting drug terbinafine. Furthermore, we identified a member of a non-canonical GET pathway client, a monotopic membrane protein squalene monooxygenase Erg1 that may be responsible for the susceptibility of Get3 mutant to lipotoxic agents.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Ivan Silbern

LAB HEAD: Prof. Dr. Henning Urlaub

PROVIDER: PXD027705 | Pride | 2022-06-09

REPOSITORIES: Pride

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			Action	DRS
	K_Bohnsack_171220_120121_Goe_L7_1.raw	Raw
	K_Bohnsack_171220_120121_Goe_L7_10.raw	Raw
	K_Bohnsack_171220_120121_Goe_L7_11.raw	Raw
	K_Bohnsack_171220_120121_Goe_L7_12.raw	Raw
	K_Bohnsack_171220_120121_Goe_L7_13.raw	Raw

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Publications

Regulated targeting of the monotopic hairpin membrane protein Erg1 requires the GET pathway.

Farkas Ákos Á Urlaub Henning H Bohnsack Katherine E KE Schwappach Blanche B

The Journal of cell biology 20220519 6

The guided entry of tail-anchored proteins (GET) pathway targets C-terminally anchored transmembrane proteins and protects cells from lipotoxicity. Here, we reveal perturbed ergosterol production in ∆get3 cells and demonstrate the sensitivity of GET pathway mutants to the sterol synthesis inhibiting drug terbinafine. Our data uncover a key enzyme of sterol synthesis, the hairpin membrane protein squalene monooxygenase (Erg1), as a non-canonical GET pathway client, thus rationalizing the lipotoxi ...[more]

PMID: 35587358

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Hepatic transcriptomics of squalene in male Apoe-deficient mice on Western diet

Project description:Background and Purpose: Squalene is the main hydrocarbon present in extra virgin olive oil and it has been reported to have anti-steatotic properties in different animal models. The aims of this study were to investigate its effects on liver transcriptomics in Male C57BL/6J Apoe-deficient mice. Experimental Approaches: Male C57BL/6J Apoe-deficient mice were fed a purified Western diet with or without squalene during 11 weeks and hepatic squalene content was assessed. Hepatic transcriptomic changes were studied and confirmed by RT-qPCR. Key Results: Squalene supplementation increased its hepatic content. The Cyp2b10 and Cyp2c55 gene expressions were significantly up-regulated by the squalene intake in all animal models, with independence of sex, sexual hormones, dietary fat content, genetic background and dose, and were strongly associated with antioxidant defense capacity and with lipid content and composition. Conclusions and Implications: hepatic squalene exerts its activity through overexpression of these cytochromes and their changes in virgin olive oil diets may be due to squalene.

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2014-06-26 | E-MTAB-1712 | biostudies-arrayexpress

A cell-free platform for characterizing and producing therapeutic bacteriophages against biohazardous and antibiotic-resistant bacteria

Project description:Bacteriophages are potent therapeutics against biohazardous bacteria that are rapidly acquiring multidrug resistance. However, routine administration of bacteriophage therapy is currently impeded by a lack of safe phage production methodologies and insufficient phage characterization. We thus developed a versatile cell-free platform for host-independent production of phages targeting gram-positive and gram-negative bacteria. A few microliters of a one-pot reaction produces effective doses of phages against potentially antibiotic-resistant bacteria such as enterohemorrhagic E. coli (EAEC) and Yersinia pestis, which also possibly pose threats as biological warfare agents. We also introduce a method for transient, non-genomic phage engineering to safely confer additional functions, such as a purification tag or bioluminescence for host detection, for only one replication cycle. Using high-resolution and time-resolved mass spectrometry, we validated the expression of 40 hypothetical proteins from two different phages (T7 and CLB-P3) and identified genes in the genome of phage T7 that express exceptionally late during phage replication. Our comprehensive methodology thus allows for accelerated reverse and forward phage engineering as well as for safe and customized production of clinical-grade therapeutic bacteriophages.

2022-07-20 | PXD019543 | Pride

Bulk RNA of IL-4 and squalene treated MDM

Project description:We performed bulk RNA-seq on squalene/M-CSF-treated macrophages to compute the fold change of genes in the squalene/M-CSF compared to M-CSF alone treated macrophages. We then looked at the distribution of the log fold changes in the skin-derived TREM2 (607 genes) and M2-like (709 genes) macrophage scRNA-seq signatures. We found that the TREM2 gene signature was more highly induced by squalene/M-CSF than the M2-like macrophage signature. In these squalene-induced macrophages, we found upregulation of several of the lipid metabolism and pro-inflammatory genes present in the skin-derived TREM2 macrophage gene signature.

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RNA-seq of Staphylococcus aureus and Staphylococcus aureus challenged with squalene compared to control growth

Project description:Squalene makes up 12 % of human skin surface lipids, however little is known about its affects on the host skin microbiome. Here we tested the effect of squalene on genetic regulation of staphylococci, showing that it profoundly affects expression virulence or colonisation determinants, and of iron uptake systems.

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Next Generation Sequencing For Quantitative Analysis Of Hepatic Transcriptomes In Response To Squalene in Swine model

Project description:Purpose: The goal of this study was to determine squalene effect on hepatic transcriptome by comparing high-throughput data for tested group vs. control group in male Large White x Landrace Large swine animals Method: Study the Hepatic mRNA profile of swine animals in response to a steatotic diet supplied with 0.5% squalene over a month by Transcriptome sequencing using DNBseq platform. Sequence reads that passed quality filters were mapped onto reference genome, followed by SNP & INDEL variants calling and Gene expression detection. Results: Squalene intake did not significantly influence single nucleotide polymorphisms. When alternative splicing events were tested, squalene administration had significant influence on splicing events including alternative 5' splicing, alternative 3' splicing , retained intron, skipped exons and mutually exclusive exons. Splicing patterns led to variety of differentially splicing genes and a variety of different isoforms from one gene. Differentially expressed genes were 18,659 in the squalene group and 18,602 in control group.

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Effect of squalene on liver transcriptome of mice lacking apolipoprotein E

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Effect of squalene on liver transcriptome of mice lacking apolipoprotein E

Project description:The hypothesis that the squalene of olive oil might influence hepatic gene expression in an apoE and sex-dependent ways was tested in mice. Gene expression was analyzed using DNA microarrays in male apoE-deficient mice that received 1 g/kg/day of squalene for 10 weeks. As initial screening of potential candidate genes involved in a differential response, only genes with remarkably modified expression (signal log2 ratio > 1.5 or < -1.5) were further considered

2016-03-30 | E-GEOD-36932 | biostudies-arrayexpress

Thraustochytrids, thraustochytrium sp.ATCC26185,LC-MSMS

Project description:This study focused on the effects of NaCl to squalene metabolism in Thraustochytrium sp. ATCC26185. The strain was cultured in different NaCl concentration medium, cells with highest specific squalene productivity were collected for proteomic analysis.

2021-06-10 | PXD021931 | Pride

The adjuvant effect of MF59 is due to the oil-in-water emulsion formulation, none of the individual components induce a comparable adjuvant effect.

Project description:Analysis of whole mouse muscle expression signature induced after 6 h by in-vivo intramuscular administration of MF59 or the individual components of MF59 (squalene oil, the surfactants Span 85 and Tween 80, either alone or in combination or the citrate buffer at the same concentrations/dose as within MF59) or PBS as control.

2013-12-30 | E-MTAB-1408 | biostudies-arrayexpress

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