Exposure to selenomethionine causes selenocysteine misincorporation and protein aggregation in Saccharomyces cerevisiae

Plateau, Pierre; Saveanu, Cosmin; Lestini, Roxane; Dauplais, Marc; Decourty, Laurence; Jacquier, Alain; Blanquet, Sylvain; Lazard, Myriam

doi:10.1038/srep44761

Cited by 47 publications

(33 citation statements)

References 55 publications

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“…Regardless of its source, as coumaric acid is important in honey bee detoxification, future experiments determining its role in metal and metalloid tolerance and whether the microbiome has a role in production of the coumaric acid-like metabolite are needed. We also observed an increase in the abundance of short-chain peptides in selenate-exposed bees, which probably indicates that protein degradation occurred as a response to treatment (92), as selenium ions have been shown to increase protein degradation in cell models (93,94). Finally, we saw higher proportional abundances of the phospholipid precursor phosphocholine (95) and of two lysophosphatidylcholines-products of oxidized phospholipids (96)-in selenate treatments as well as MSEA results that indicated overexpression of phospholipid biosynthesis metabolites in both treatments.…”

Section: Toxicants Affect the Bee Microbiome And Metabolomementioning

confidence: 63%

Cadmium and Selenate Exposure Affects the Honey Bee Microbiome and Metabolome, and Bee-Associated Bacteria Show Potential for Bioaccumulation

Rothman

Leger

Kirkwood

et al. 2019

Appl Environ Microbiol

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Honey bees are important insect pollinators used heavily in agriculture and can be found in diverse environments. Bees may encounter toxicants such as cadmium and selenate by foraging on plants growing in contaminated areas, which can result in negative health effects. Honey bees are known to have a simple and consistent microbiome that conveys many benefits to the host, and toxicant exposure may impact this symbiotic microbial community. We used 16S rRNA gene sequencing to assay the effects that sublethal cadmium and selenate treatments had over 7 days and found that both treatments significantly but subtly altered the composition of the bee microbiome. Next, we exposed bees to cadmium and selenate and then used untargeted liquid chromatography-mass spectrometry (LC-MS) metabolomics to show that chemical exposure changed the bees' metabolite profiles and that compounds which may be involved in detoxification, proteolysis, and lipolysis were more abundant in treatments. Finally, we exposed several strains of bee-associated bacteria in liquid culture and found that each strain removed cadmium from its medium but that only Lactobacillus Firm-5 microbes assimilated selenate, indicating the possibility that these microbes may reduce the metal and metalloid burden on their host. Overall, our report shows that metal and metalloid exposure can affect the honey bee microbiome and metabolome and that strains of bee-associated bacteria can bioaccumulate these toxicants. IMPORTANCE Bees are important insect pollinators that may encounter environmental pollution when foraging upon plants grown in contaminated areas. Despite the pervasiveness of pollution, little is known about the effects of these toxicants on honey bee metabolism and their symbiotic microbiomes. Here, we investigated the impact of selenate and cadmium exposure on the gut microbiome and metabolome of honey bees. We found that exposure to these chemicals subtly altered the overall composition of the bees' microbiome and metabolome and that exposure to toxicants may negatively impact both host and microbe. As the microbiome of animals can reduce mortality upon metal or metalloid challenge, we grew bee-associated bacteria in media spiked with selenate or cadmium. We show that some bacteria can remove these toxicants from their media in vitro and suggest that bacteria may reduce metal burden in their hosts.

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Section: Toxicants Affect the Bee Microbiome And Metabolomementioning

confidence: 63%

Cadmium and Selenate Exposure Affects the Honey Bee Microbiome and Metabolome, and Bee-Associated Bacteria Show Potential for Bioaccumulation

Rothman

Leger

Kirkwood

et al. 2019

Appl Environ Microbiol

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“…Bioavailability, toxicity and antioxidant activity of selenium depends on its chemical form. Selenomethionine is the most widely used form of Se in food and supplements, though it has less toxicity and excellent bioavailability but some reports show that its excessive use can lead to toxic effects [46]. Therefore, instead of selenomethionine Nano-Se can be used as its effect on upregulating glutathione peroxidase and thioredoxin reductase is comparable with selenomethionine with much lower toxicity [38].…”

Section: Therapeutic Efficacy Of Se Based Nanoparticlesmentioning

confidence: 99%

Therapeutic Potential of Selenium Nanoparticles

Majeed¹,

Zafar²,

Bhatti³

et al. 2018

J Nanomed Nanotechnol

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“…As these studies indicated, SeMet toxicity appears to come from two different mechanisms: SeMet likely generates reactive species, resulting in DNA damage 28 , consistent with the observation that SeMet could effectively inhibit the insect cell growth 17 . Alternatively, SeMet toxicity results from its metabolites, and a random incorporation of which promotes protein aggregation -SeMet causes proteotoxic stress resulting in cell death 29 . How does baculovirus infection reduce SeMet toxicity?…”

Section: Possible Mechanism Of Semet Toxicitymentioning

confidence: 99%

A practical method for efficient and optimal production of selenomethionine-labeled recombinant protein complexes in the insect cells

Wenzel

Imasaki

Takagi

2018

Preprint

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The use of Selenomethionine (SeMet) incorporated protein crystals for single or multiwavelength anomalous diffraction (SAD or MAD) to facilitate phasing has become almost synonymous with modern X-ray crystallography. The anomalous signals from SeMets can be used for phasing as well as sequence markers for subsequent model building. The production of large quantities of SeMet incorporated recombinant proteins is relatively straightforward when expressed in E. coli. In contrast, production of SeMet substituted recombinant proteins expressed in the insect cells is not as robust due to the toxicity of SeMet in eukaryotic systems. Previous protocols for SeMet-incorporation in the insect cells are laborious, and more suited for secreted proteins. In addition, these protocols have generally not addressed the SeMet toxicity issue, and typically result in low recovery of the labeled proteins. Here we report that SeMet toxicity can be circumvented by fully infecting insect cells with baculovirus. Quantitatively controlling infection levels using our Titer Estimation of Quality Control (TEQC) method allows for incorporation of substantial amounts of SeMet, resulting in an efficient and optimal production of labeled recombinant protein complexes. With the method described here, we were able to consistently reach incorporation levels of about 75% and protein yield of 60-90% compared to native protein expression. Keywords:Seleno-methionine (SeMet), baculovirus expression vector system, Sf9 and Hi5 insect cells, TEQC method, baculovirus infectivity, estimated multiplicity of infectivity (eMOI), the Mediator Head module, SeM-TEQC method Abbreviations and symbols IntroductionSingle or multi-wavelength anomalous diffraction (SAD or MAD) phasing by utilizing Seleno-methionine (SeMet) labeled proteins or protein complexes is almost synonymous with modern X-ray crystallography 1,2 . This method has drastically reduced the need for preparation of heavy atom derivatives, which is one of the most timeconsuming steps in X-ray crystallography. The anomalous signal from SeMet can be used for phasing as well as for identifying methionine positions within the electron density map, thereby aiding model building in low resolution maps 3,4 .Production of SeMet-labeled proteins in E. coli for X-ray crystallography is well established and robust 1 . Protocols exist for yeast Saccharomyces cerevisiae 3 , yeast Pichia pastoris 5 , the insect cells 6,7 as well as mammalian cells 8 . While SeMet incorporation routinely achieves 100% for proteins expressed in E. coli, expression in eukaryotic cells is much more variable, with SeMet incorporation ranging from 50 % to 90 %, depending on the host cells and proteins being produced 5,7-10 . The baculovirus expression system (BEVS) is an excellent tool for expressing recombinant proteins in insect cells. It is considered an attractive option for proteins as well as protein complexes and has been widely used for structural and functional studies [11][12][13] . For production of SeMet-labelled proteins using ...

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Exposure to selenomethionine causes selenocysteine misincorporation and protein aggregation in Saccharomyces cerevisiae

Cited by 47 publications

References 55 publications

Cadmium and Selenate Exposure Affects the Honey Bee Microbiome and Metabolome, and Bee-Associated Bacteria Show Potential for Bioaccumulation

Cadmium and Selenate Exposure Affects the Honey Bee Microbiome and Metabolome, and Bee-Associated Bacteria Show Potential for Bioaccumulation

Therapeutic Potential of Selenium Nanoparticles

A practical method for efficient and optimal production of selenomethionine-labeled recombinant protein complexes in the insect cells

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