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1.
J Biol Chem ; 300(1): 105562, 2024 Jan.
Article En | MEDLINE | ID: mdl-38097189

Extensive epigenetic reprogramming occurs during preimplantation embryonic development. However, the impact of DNA methylation in plateau yak preimplantation embryos and how epigenetic reprogramming contributes to transcriptional regulatory networks are unclear. In this study, we quantified gene expression and DNA methylation in oocytes and a series of yak embryos at different developmental stages and at single-cell resolution using single-cell bisulfite-sequencing and RNA-seq. We characterized embryonic genome activation and maternal transcript degradation and mapped epigenetic reprogramming events critical for embryonic development. Through cross-species transcriptome analysis, we identified 31 conserved maternal hub genes and 39 conserved zygotic hub genes, including SIN3A, PRC1, HDAC1/2, and HSPD1. Notably, by combining single-cell DNA methylation and transcriptome analysis, we identified 43 candidate methylation driver genes, such as AURKA, NUSAP1, CENPF, and PLK1, that may be associated with embryonic development. Finally, using functional approaches, we further determined that the epigenetic modifications associated with the histone deacetylases HDAC1/2 are essential for embryonic development and that the deubiquitinating enzyme USP7 may affect embryonic development by regulating DNA methylation. Our data represent an extensive resource on the transcriptional dynamics of yak embryonic development and DNA methylation remodeling, and provide new insights into strategies for the conservation of germplasm resources, as well as a better understanding of mammalian early embryonic development that can be applied to investigate the causes of early developmental disorders.


Blastocyst , DNA Methylation , Embryo, Mammalian , Embryonic Development , Gene Expression Regulation, Developmental , Single-Cell Gene Expression Analysis , Sulfites , Animals , Cattle , Female , Pregnancy , Blastocyst/metabolism , Embryonic Development/genetics , Epigenesis, Genetic , Gene Expression Profiling , Histone Deacetylase 1/metabolism , Histone Deacetylase 2/metabolism , Sulfites/metabolism , Ubiquitin-Specific Peptidase 7/metabolism , Embryo, Mammalian/embryology , Embryo, Mammalian/enzymology
2.
Methods Enzymol ; 692: 23-38, 2023.
Article En | MEDLINE | ID: mdl-37925181

Posttranscriptional RNA modifications occur in almost all types of RNA in all life forms. As an abundant RNA modification in mammals, pseudouridine (Ψ) regulates diverse biological functions of different RNA species such as ribosomal RNA (rRNA), transfer RNA (tRNA), small nuclear RNA (snRNA), etc. However, the functional investigation of mRNA pseudouridine (Ψ) has been hampered by the lack of a quantitative method that can efficiently map Ψ transcriptome-wide. We developed bisulfite-induced deletion sequencing (BID-seq), with the optimized bisulfite-based chemical reaction to convert pseudouridine selectively and completely into Ψ-BS adduct without cytosine deamination. The Ψ-BS adduct can be further read out as deletion signatures during reverse transcription. The deletion ratios induced by Ψ sites were used for estimating the modification stoichiometry at each modified site. BID-seq starts with 10-20 ng polyA+ RNA and detects thousands of mRNA Ψ sites with stoichiometry information in cell lines and tissues. We uncovered consensus motifs for Ψ in mammalian mRNA and assigned specific 'writer' proteins to individual Ψ deposition. BID-seq also confirmed the presence of Ψ within stop codons of mammalian mRNA. BID-seq set the stage for future investigations of Ψ functions in diverse biological processes.


Pseudouridine , RNA , Animals , RNA, Messenger/genetics , RNA, Messenger/metabolism , Pseudouridine/metabolism , RNA/metabolism , Sulfites/metabolism , RNA, Ribosomal/metabolism , RNA, Transfer/metabolism , RNA Processing, Post-Transcriptional , Mammals/genetics , Mammals/metabolism
3.
FEMS Microbiol Rev ; 47(5)2023 09 05.
Article En | MEDLINE | ID: mdl-37796897

Sulfate/sulfite-reducing microorganisms (SRM) are ubiquitous in nature, driving the global sulfur cycle. A hallmark of SRM is the dissimilatory sulfite reductase encoded by the genes dsrAB. Based on analysis of 950 mainly metagenome-derived dsrAB-carrying genomes, we redefine the global diversity of microorganisms with the potential for dissimilatory sulfate/sulfite reduction and uncover genetic repertoires that challenge earlier generalizations regarding their mode of energy metabolism. We show: (i) 19 out of 23 bacterial and 2 out of 4 archaeal phyla harbor uncharacterized SRM, (ii) four phyla including the Desulfobacterota harbor microorganisms with the genetic potential to switch between sulfate/sulfite reduction and sulfur oxidation, and (iii) the combination as well as presence/absence of different dsrAB-types, dsrL-types and dsrD provides guidance on the inferred direction of dissimilatory sulfur metabolism. We further provide an updated dsrAB database including > 60% taxonomically resolved, uncultured family-level lineages and recommendations on existing dsrAB-targeted primers for environmental surveys. Our work summarizes insights into the inferred ecophysiology of newly discovered SRM, puts SRM diversity into context of the major recent changes in bacterial and archaeal taxonomy, and provides an up-to-date framework to study SRM in a global context.


Archaea , Bacteria , Oxidation-Reduction , Bacteria/metabolism , Archaea/metabolism , Sulfates/metabolism , Sulfites/metabolism , Sulfur/metabolism , Phylogeny
4.
J Biol Chem ; 299(8): 105010, 2023 08.
Article En | MEDLINE | ID: mdl-37414148

The obligately anaerobic sulfite-reducing bacterium Bilophila wadsworthia is a common human pathobiont inhabiting the distal intestinal tract. It has a unique ability to utilize a diverse range of food- and host-derived sulfonates to generate sulfite as a terminal electron acceptor (TEA) for anaerobic respiration, converting the sulfonate sulfur to H2S, implicated in inflammatory conditions and colon cancer. The biochemical pathways involved in the metabolism of the C2 sulfonates isethionate and taurine by B. wadsworthia were recently reported. However, its mechanism for metabolizing sulfoacetate, another prevalent C2 sulfonate, remained unknown. Here, we report bioinformatics investigations and in vitro biochemical assays that uncover the molecular basis for the utilization of sulfoacetate as a source of TEA (STEA) for B. wadsworthia, involving conversion to sulfoacetyl-CoA by an ADP-forming sulfoacetate-CoA ligase (SauCD), and stepwise reduction to isethionate by NAD(P)H-dependent enzymes sulfoacetaldehyde dehydrogenase (SauS) and sulfoacetaldehyde reductase (TauF). Isethionate is then cleaved by the O2-sensitive isethionate sulfolyase (IseG), releasing sulfite for dissimilatory reduction to H2S. Sulfoacetate in different environments originates from anthropogenic sources such as detergents, and natural sources such as bacterial metabolism of the highly abundant organosulfonates sulfoquinovose and taurine. Identification of enzymes for anaerobic degradation of this relatively inert and electron-deficient C2 sulfonate provides further insights into sulfur recycling in the anaerobic biosphere, including the human gut microbiome.


Bilophila , Humans , Alkanesulfonates/metabolism , Bilophila/metabolism , Sulfites/metabolism , Sulfur/metabolism , Taurine/metabolism , Gastrointestinal Microbiome
5.
Int J Biol Macromol ; 240: 124428, 2023 Jun 15.
Article En | MEDLINE | ID: mdl-37062383

Bilophila wadsworthia is one of the prominent sources of hydrogen sulfide (H2S) production in appendices, excessive levels of which can result in a weaker colonic mucus barrier, inflammatory bowel disease, and colorectal cancer. Isethionate sulfite-lyase (IslA) enzyme catalyzes H2S production by cleaving CS bond in isethionate, producing acetaldehyde and sulfite. In this study, we aimed to identify potential substrate antagonists for IsIA using a structure-based drug design. Initially, pharmacophore-based computational screening of the ZINC20 database yielded 66 hits that were subjected to molecular docking targeting the isethionate binding site of IsIA. Based on striking docking scores, nine compounds showed strong interaction with critical IsIA residues (Arg189, Gln193, Glu470, Cys468, and Arg678), drug-like features, appropriate adsorption, metabolism, excretion, and excretion profile with non-toxicity. Molecular dynamics simulations uncovered the significant impact of binding the compounds on protein conformational dynamics. Finally, binding free energies revealed substantial binding affinity (ranging from -35.23 to -53.88 kcal/mol) of compounds (ZINC913876497, ZINC913856647, ZINC914263733, ZINC914137795, ZINC915757996, ZINC914357083, ZINC913934833, ZINC9143362047, and ZINC913854740) for IsIA. The compounds proposed herein through a multi-faceted computational strategy can be experimentally validated as potential substrate antagonists of B. wadsworthia's IsIA for developing new medications to curb gut-associated illness in the future.


Bilophila , Lyases , Molecular Docking Simulation , Bilophila/metabolism , Lyases/metabolism , Molecular Dynamics Simulation , Sulfites/metabolism , Ligands
6.
Microbiol Spectr ; 11(3): e0132623, 2023 06 15.
Article En | MEDLINE | ID: mdl-37098949

Selenium (Se) is a micronutrient in most eukaryotes, and Se-enriched yeast is the most common selenium supplement. However, selenium metabolism and transport in yeast have remained unclear, greatly hindering the application of this element. To explore the latent selenium transport and metabolism mechanisms, we performed adaptive laboratory evolution under the selective pressure of sodium selenite and successfully obtained selenium-tolerant yeast strains. Mutations in the sulfite transporter gene ssu1 and its transcription factor gene fzf1 were found to be responsible for the tolerance generated in the evolved strains, and the selenium efflux process mediated by ssu1 was identified in this study. Moreover, we found that selenite is a competitive substrate for sulfite during the efflux process mediated by ssu1, and the expression of ssu1 is induced by selenite rather than sulfite. Based on the deletion of ssu1, we increased the intracellular selenomethionine content in Se-enriched yeast. This work confirms the existence of the selenium efflux process, and our findings may benefit the optimization of Se-enriched yeast production in the future. IMPORTANCE Selenium is an essential micronutrient for mammals, and its deficiency severely threatens human health. Yeast is the model organism for studying the biological role of selenium, and Se-enriched yeast is the most popular selenium supplement to solve Se deficiency. The cognition of selenium accumulation in yeast always focuses on the reduction process. Little is known about selenium transport, especially selenium efflux, which may play a crucial part in selenium metabolism. The significance of our research is in determining the selenium efflux process in Saccharomyces cerevisiae, which will greatly enhance our knowledge of selenium tolerance and transport, facilitating the production of Se-enriched yeast. Moreover, our research further advances the understanding of the relationship between selenium and sulfur in transport.


Saccharomyces cerevisiae , Selenium , Humans , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Selenium/metabolism , Selenium/pharmacology , Selenomethionine/metabolism , Sulfites/metabolism , Selenious Acid/metabolism
7.
Environ Sci Technol ; 57(6): 2371-2379, 2023 02 14.
Article En | MEDLINE | ID: mdl-36734488

Microbial transformation of selenite [Se(IV)] to elemental selenium nanoparticles (SeNPs) is known to be an important process for removing toxic soluble selenium (Se) oxyanions and recovery of Se from the environment as valuable nanoparticles. However, the mechanism of selenite uptake by microorganisms, the first step through which Se exerts its cellular function, remains not well studied. In this study, the effects of selenite concentration, time, pH, metabolic inhibitors, and anionic analogues on selenite uptake in Rahnella aquatilis HX2 were investigated. Selenite uptake by R. aquatilis HX2 was concentration- and time-dependent, and its transport activity was significantly dependent on pH. In addition, selenite uptake in R. aquatilis HX2 was significantly inhibited by the aquaporin inhibitor AgNO3 and sulfite (SO32-), and partially inhibited by carbonyl cyanide m-chlorophenyl hydrazone (CCCP) and 2,4-dinitrophenol (2,4-DNP) treatments. Three mutants with in-frame deletions of aqpZ, glpF, and nhaA genes were constructed. The transport assay showed that the water channel protein AqpZ, and not GlpF, was a key channel of selenite uptake by R. aquatilis HX2, and sulfite and selenite had a common uptake pathway. In addition, the Na+/H+ antiporter NhaA is also involved in selenite uptake in R. aquatilis HX2.


Rahnella , Selenium , Selenium/chemistry , Selenium/metabolism , Rahnella/genetics , Rahnella/metabolism , Selenious Acid/pharmacology , Selenious Acid/metabolism , Ions/metabolism , Sulfites/metabolism
8.
Anal Chem ; 95(2): 1556-1565, 2023 01 17.
Article En | MEDLINE | ID: mdl-36563112

DNA methylation (5-methylcytosine, 5mC) is the most important epigenetic modification in mammals. Deciphering the roles of 5mC relies on the quantitative detection of 5mC at the single-base resolution. Bisulfite sequencing (BS-seq) is the most often employed technique for mapping 5mC in DNA. However, bisulfite treatment may cause serious degradation of input DNA due to the harsh reaction conditions. Here, we engineered the human apolipoprotein B mRNA-editing catalytic polypeptide-like 3C (A3C) protein to endow the engineered A3C (eA3C) protein with differential deamination activity toward cytosine and 5mC. By the virtue of the unique property of eA3C, we proposed an engineered A3C sequencing (EAC-seq) method for the bisulfite-free and quantitative mapping of 5mC in DNA at the single-base resolution. In EAC-seq, the eA3C protein can deaminate C but not 5mC, which is employed to differentiate C and 5mC in sequencing. Using the EAC-seq method, we quantitatively detected 5mC in genomic DNA of lung cancer tissue. In contrast to the harsh reaction conditions of BS-seq, which could lead to significant degradation of DNA, the whole procedure of EAC-seq is carried out under mild conditions, thereby preventing DNA damage. Taken together, the EAC-seq approach is bisulfite-free and straightforward, making it an invaluable tool for the quantitative detection of 5mC in limited DNA at the single-base resolution.


5-Methylcytosine , Cytidine Deaminase , DNA Methylation , Humans , 5-Methylcytosine/metabolism , Cytidine Deaminase/genetics , Cytidine Deaminase/metabolism , Cytosine , DNA/genetics , DNA/metabolism , Epigenesis, Genetic , Sequence Analysis, DNA/methods , Sulfites/metabolism
9.
BMC Plant Biol ; 22(1): 491, 2022 Oct 18.
Article En | MEDLINE | ID: mdl-36253724

BACKGROUND: ATP sulfurylase (ATPS) is a crucial enzyme for the selenate assimilation pathway in plants. RESULTS: In this study, genome-wide and comparative analyses of ATPS in Cardamine hupingshanensis, including sequence and structural analyses, were performed. The expression of ChATPS gene family members in C. hupingshanensis under selenium (Se) stress was also investigated, and our results suggest that ChATPS1-2 play key roles in the response to Se stress. Nine ATPS genes were found from C. hupingshanensis, which share highly conserved sequences with ATPS from Arabidopsis thaliana. In addition, we performed molecular docking of ATP sulfurylase in complex with compounds ATP, selenate, selenite, sulfate, and sulfite. ChAPS3-1 was found to have stronger binding energies with all compounds tested. Among these complexes, amino acid residues Arg, Gly, Ser, Glu, and Asn were commonly present. CONCLUSION: Our study reveals the molecular mechanism of C. hupingshanensis ATP sulfurylase interacting with selenate, which is essential for understanding selenium assimilation. This information will guide further studies on the function of the ChATPS gene family in the selenium stress response and lay the foundation for the selenium metabolic pathway in higher plants.


Arabidopsis , Cardamine , Selenium , Adenosine Triphosphate , Amino Acids/metabolism , Arabidopsis/metabolism , Cardamine/metabolism , Molecular Docking Simulation , Selenic Acid , Selenious Acid/metabolism , Selenium/metabolism , Sulfate Adenylyltransferase/chemistry , Sulfate Adenylyltransferase/genetics , Sulfate Adenylyltransferase/metabolism , Sulfates/metabolism , Sulfites/metabolism
10.
BMC Bioinformatics ; 23(1): 383, 2022 Sep 19.
Article En | MEDLINE | ID: mdl-36123620

BACKGROUND: DNA methylation is an important epigenetic modification that is known to regulate gene expression. Whole-genome bisulfite sequencing (WGBS) is a powerful method for studying cytosine methylation in a whole genome. However, it is difficult to obtain methylation profiles using the WGBS raw reads and is necessary to be proficient in all types of bioinformatic tools for the study of DNA methylation. In addition, recent end-to-end pipelines for DNA methylation analyses are not sufficient for addressing those difficulties. RESULTS: Here we present msPIPE, a pipeline for DNA methylation analyses with WGBS data seamlessly connecting all the required tasks ranging from data pre-processing to multiple downstream DNA methylation analyses. The msPIPE can generate various methylation profiles to analyze methylation patterns in the given sample, including statistical summaries and methylation levels. Also, the methylation levels in the functional regions of a genome are computed with proper annotation. The results of methylation profiles, hypomethylation, and differential methylation analysis are plotted in publication-quality figures. The msPIPE can be easily and conveniently used with a Docker image, which includes all dependent packages and software related to DNA methylation analyses. CONCLUSION: msPIPE is a new end-to-end pipeline designed for methylation calling, profiling, and various types of downstream DNA methylation analyses, leading to the creation of publication-quality figures. msPIPE allows researchers to process and analyze the WGBS data in an easy and convenient way. It is available at https://github.com/jkimlab/msPIPE and https://hub.docker.com/r/jkimlab/mspipe .


Cytosine , Sulfites , Sequence Analysis, DNA/methods , Sulfites/metabolism , Whole Genome Sequencing/methods
11.
J Hazard Mater ; 439: 129685, 2022 10 05.
Article En | MEDLINE | ID: mdl-36104911

Sulfur dioxide (SO2) and its derivatives are known to be hazardous but their common application in food, especially the grape industry, is conditionally allowed. Potential hazards to consumers and the environment could occur upon the control-lost SO2 during grape logistics and storage. Researchers have usually focused on the anti-pathogen role of SO2 whereas limited efforts were conducted on the sulfur (S) absorption, assimilation patterns, and sulfite detoxification. In this study, short-term, room-temperature, and SO2-stored grapes were investigated, whose S flux of various forms was quantified through an estimation model. Accordingly, the additional accumulated S (0.50-0.86%) in pulps from atmospheric SO2 was considered mainly through rachis transport compared to across skin surfaces and the usage arrangement of the absorbed S was included. The first quantitative evidence of induced S assimilation under SO2 was also provided, which challenged the previous knowledge. In addition, sulfite oxidase and reductase (SiO and SiR) played major roles in sulfite detoxification, being effectively stimulated at multiple levels. The induced S metabolism associated with enhanced reactive oxygen species (ROS) scavenging capacity and alleviated senescence contributed to quality maintenance. Overall, these findings provide novel insights and are valuable supports for developing SO2-controlling strategies to avoid potential hazards.


Sulfite Oxidase , Vitis , Sulfite Oxidase/metabolism , Sulfites/metabolism , Sulfur/metabolism , Sulfur Dioxide
12.
Sci Total Environ ; 852: 158203, 2022 Dec 15.
Article En | MEDLINE | ID: mdl-36044953

Sulfur (S) is a crucial component in the environment and living organisms. This work is the first attempt to provide an overview and critical discussion on the roles, mechanisms, and environmental applications of sulfur-oxidizing bacteria (SOB). The findings reveal that key enzymes of SOB embarked on oxidation of sulfide, sulfite, thiosulfate, and elemental S. Conversion of reduced S compounds was oxidatively catalyzed by various enzymes (e.g. sulfide: quinone oxidoreductase, flavocytochrome c-sulfide dehydrogenase, dissimilatory sulfite reductase, heterodisulfide reductase-like proteins). Environmental applications of SOB discussed include detoxifying hydrogen sulfide, soil bioremediation, and wastewater treatment. SOB producing S0 engaged in biological S soil amendments (e.g. saline-alkali soil remediation, the oxidation of sulfide-bearing minerals). Biotreatment of H2S using SOB occurred under both aerobic and anaerobic conditions. Sulfide, nitrate, and sulfamethoxazole were removed through SOB suspension cultures and S0-based carriers. Finally, this work presented future perspectives on SOB development, including S0 recovery, SOB enrichment, field measurement and identification of sulfur compounds, and the development of mathematical simulation.


Hydrogen Sulfide , Biodegradation, Environmental , Hydrogensulfite Reductase/metabolism , Thiosulfates , Nitrates/metabolism , Sulfur/metabolism , Bacteria/metabolism , Oxidation-Reduction , Oxidoreductases/metabolism , Sulfides/metabolism , Soil , Sulfamethoxazole/metabolism , Sulfites/metabolism , Alkalies , Quinones
13.
J Hazard Mater ; 440: 129703, 2022 10 15.
Article En | MEDLINE | ID: mdl-35963088

The removal of environmental pollutants is important for a sustainable ecosystem and human health. Shewanella oneidensis (S. oneidensis) has diverse electron transfer pathways and can use a variety of contaminants as electron acceptors or electron donors. This paper reviews S. oneidensis's function in removing environmental pollutants, including heavy metals, inorganic non-metallic ions (INMIs), and toxic organic pollutants. S. oneidensis can mineralize o-xylene (OX), phenanthrene (PHE), and pyridine (Py) as electron donors, and also reduce azo dyes, nitro aromatic compounds (NACs), heavy metals, and iodate by extracellular electron transfer (EET). For azo dyes, NACs, Cr(VI), nitrite, nitrate, thiosulfate, and sulfite that can cross the membrane, S. oneidensis transfers electrons to intracellular reductases to catalyze their reduction. However, most organic pollutants cannot be directly degraded by S. oneidensis, but S. oneidensis can remove these pollutants by self-synthesizing catalysts or photocatalysts, constructing bio-photocatalytic systems, driving Fenton reactions, forming microbial consortia, and genetic engineering. However, the industrial-scale application of S. oneidensis is insufficient. Future research on the metabolism of S. oneidensis and interfacial reactions with other materials needs to be deepened, and large-scale reactors should be developed that can be used for practical engineering applications.


Environmental Pollutants , Phenanthrenes , Shewanella , Azo Compounds/metabolism , Ecosystem , Electron Transport , Environmental Pollutants/metabolism , Iodates/metabolism , Nitrates/metabolism , Nitrites/metabolism , Oxidation-Reduction , Oxidoreductases/metabolism , Phenanthrenes/metabolism , Pyridines/metabolism , Shewanella/metabolism , Sulfites/metabolism , Thiosulfates/metabolism
14.
Nat Commun ; 13(1): 4110, 2022 07 15.
Article En | MEDLINE | ID: mdl-35840579

The Terrestrial Miscellaneous Euryarchaeota Group has been identified in various environments, and the single genome investigated thus far suggests that these archaea are anaerobic sulfite reducers. We assemble 35 new genomes from this group that, based on genome analysis, appear to possess aerobic and facultative anaerobic lifestyles and may oxidise rather than reduce sulfite. We propose naming this order (representing 16 genera) "Lutacidiplasmatales" due to their occurrence in various acidic environments and placement within the phylum Thermoplasmatota. Phylum-level analysis reveals that Thermoplasmatota evolution had been punctuated by several periods of high levels of novel gene family acquisition. Several essential metabolisms, such as aerobic respiration and acid tolerance, were likely acquired independently by divergent lineages through convergent evolution rather than inherited from a common ancestor. Ultimately, this study describes the terrestrially prevalent Lutacidiciplasmatales and highlights convergent evolution as an important driving force in the evolution of archaeal lineages.


Archaeal Proteins , Euryarchaeota , Archaea/metabolism , Archaeal Proteins/genetics , Archaeal Proteins/metabolism , Euryarchaeota/genetics , Evolution, Molecular , Genome, Archaeal/genetics , Phylogeny , Sulfites/metabolism
15.
Proc Natl Acad Sci U S A ; 119(11): e2118002119, 2022 03 15.
Article En | MEDLINE | ID: mdl-35271389

SignificanceYeiE has been identified as a master virulence factor of Cronobacter sakazakii. In this study, we determined the crystal structures of the regulatory domain of YeiE in complex with its physiological ligand sulfite ion (SO32-). The structure provides the basis for the molecular mechanisms for sulfite sensing and the ligand-dependent conformational changes of the regulatory domain. The genes under the control of YeiE in response to sulfite were investigated to reveal the functional roles of YeiE in the sulfite tolerance of the bacteria. We propose the molecular mechanism underlying the ability of gram-negative pathogens to defend against the innate immune response involving sulfite, thus providing a strategy to control the pathogenesis of bacteria.


Bacterial Proteins , Cronobacter sakazakii , Stress, Physiological , Sulfites , Transcription Factors , Virulence Factors , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Cronobacter sakazakii/genetics , Cronobacter sakazakii/metabolism , Cronobacter sakazakii/pathogenicity , Crystallization , Ligands , Protein Domains , Sulfites/metabolism , Transcription Factors/chemistry , Transcription Factors/genetics , Virulence Factors/chemistry , Virulence Factors/genetics
16.
Cells ; 11(4)2022 02 11.
Article En | MEDLINE | ID: mdl-35203287

Saccharomyces uvarum is one of the few fermentative species that can be used in winemaking, but its weak sulfite tolerance is the main reason for its further use. Previous studies have shown that the expression of the methionine synthase gene (MET4) is upregulated in FZF1 (a gene encoding a putative zinc finger protein, which is a positive regulator of the transcription of the cytosolic sulfotransferase gene SSU1) overexpression transformant strains, but its exact function is unknown. To gain insight into the function of the MET4 gene, in this study, a MET4 overexpression vector was constructed and transformed into S. uvarum strain A9. The MET4 transformants showed a 20 mM increase in sulfite tolerance compared to the starting strain. Ninety-two differential genes were found in the transcriptome of A9-MET4 compared to the A9 strain, of which 90 were upregulated, and two were downregulated. The results of RT-qPCR analyses confirmed that the expression of the HOMoserine requiring gene (HOM3) in the sulfate assimilation pathway and some fermentation-stress-related genes were upregulated in the transformants. The overexpression of the MET4 gene resulted in a significant increase in sulfite tolerance, the upregulation of fermentation-stress-related gene expression, and significant changes in the transcriptome profile of the S. uvarum strain.


Basic-Leucine Zipper Transcription Factors , Fungal Proteins , Saccharomyces , Sulfites , Basic-Leucine Zipper Transcription Factors/genetics , Basic-Leucine Zipper Transcription Factors/metabolism , Fungal Proteins/genetics , Fungal Proteins/metabolism , Saccharomyces/genetics , Saccharomyces/metabolism , Sulfites/metabolism , Up-Regulation
17.
Methods Mol Biol ; 2443: 451-463, 2022.
Article En | MEDLINE | ID: mdl-35037220

The mechanism of the addition of a methyl group to cytosine has been identified as one of several heritable epigenetic mechanisms. In plants, DNA methylation is involved in mediating response to stress, plant development, polyploidy, and domestication through regulation of gene expression. The correlation of epigenetic variation to phenotypic traits expands our understanding toward plant evolution, and provides new source for targeted manipulation in crop improvement. To address the increasing interest to map methylation landscape in plant species, this chapter describes methods to analyze bisulfite sequencing data and identify epigenetic variation between samples. We also detailed guidelines to highlight possible optimizations, as well as ways to tailor parameters according to data and biological variability.


Cytosine , Sulfites , Cytosine/metabolism , DNA Methylation , Epigenesis, Genetic , Sulfites/metabolism
18.
J Exp Bot ; 73(8): 2525-2539, 2022 04 18.
Article En | MEDLINE | ID: mdl-35084469

The roles of cytosolic O-acetylserine-(thiol)-lyase A (OASTLA), chloroplastic OASTLB, and mitochondrial OASTLC in plant selenate resistance were studied in Arabidopsis. Impairment in OASTLA and OASTLB resulted in reduced biomass, chlorophyll and soluble protein content compared with selenate-treated OASTLC-impaired and wild-type plants. The generally lower total selenium (Se), protein-Se, organic-sulfur and protein-sulfur (S) content in oastlA and oastlB compared with wild-type and oastlC leaves indicated that Se accumulation was not the main cause for the stress symptoms in these mutants. Notably, the application of selenate positively induced S-starvation markers and the OASTLs, followed by increased sulfite reductase, sulfite oxidase activities, and increased sulfite and sulfide concentrations. Taken together, our results indicate a futile anabolic S-starvation response that resulted in lower glutathione and increased oxidative stress symptoms in oastlA and oastlB mutants. In-gel assays of l-cysteine and l-seleno-cysteine, desulfhydrase activities revealed that two of the three OASTL activity bands in each of the oastl single mutants were enhanced in response to selenate, whereas the impaired proteins exhibited a missing activity band. The absence of differently migrated activity bands in each of the three oastl mutants indicates that these OASTLs are major components of desulfhydrase activity, degrading l-cysteine and l-seleno-cysteine in Arabidopsis.


Arabidopsis , Lyases , Selenium , Arabidopsis/metabolism , Carbon-Oxygen Lyases/metabolism , Cysteine/metabolism , Lyases/metabolism , Selenic Acid , Selenium/metabolism , Serine/analogs & derivatives , Sulfhydryl Compounds/metabolism , Sulfites/metabolism , Sulfur/metabolism
19.
Chemistry ; 28(23): e202104342, 2022 Apr 22.
Article En | MEDLINE | ID: mdl-35080290

Sulfite is a potent toxic substance causing harm to multi-organ in human. Despite toxicity, it is widely used as preservative, anti-browning and anti-oxidant in foods, beverages, and pharmaceuticals, which cause easy admission of sulfite in human. Sulfite is also produced endogenously during the catabolism of cysteine and methionine. In vivo, the serum sulfite level at physiological range is strictly maintained by a molybdenum dependent sulfite oxidase (SO), which catalyzes sulfite to sulfate oxidation via a two-electron oxidation pathway. The loss of SO activity causes high serum sulfite level that fosters several diseases, including asthma, neurological dysfunction, birth defects, and heart diseases. The cytotoxicity of (bi)sulfite is implicated as sulfite radicals, which are generated by mainly heme-peroxidases via a one-electron oxidation pathway. On the other hand, the toxic sulfite radicals are neutralized to sulfite by heme-globins. The enzymatic reduction of sulfite to sulfide is catalyzed by sulfite reductase, which contains an unusual metal cofactor, siroheme-[4Fe4S]-cluster. Overall, the interaction of sulfite with various metalloproteins in vivo is a close relation with human health. Therefore, this review describes the metabolic conversion of (bi)sulfite to sulfate, sulfite radical or sulfide via oxidation or reduction pathways by various metalloproteins (specially SOs, peroxidases, heme-globins, and sulfite reductases), and the potential applications of sulfite in biosensors/biofuel cells, anti-browning, and advance oxidation process.


Metalloproteins , Globins , Heme , Humans , Hydrogen , Metalloproteins/metabolism , Oxidoreductases Acting on Sulfur Group Donors/metabolism , Peroxidases , Sulfates , Sulfides , Sulfites/metabolism
20.
J Sci Food Agric ; 102(3): 1174-1184, 2022 Feb.
Article En | MEDLINE | ID: mdl-34338316

BACKGROUND: The commercial preservation of table grapes largely depends on the application of sulfur dioxide (SO2 ). However, little is known about whether SO2 participates in sulfur metabolism to improve the postharvest quality of table grapes. In this study, the contents of sulfur-containing compounds, activities of enzymes, and expression of genes involved in sulfur metabolism in table grapes (Vitis vinifera cv. Thompson Seedless) were evaluated. RESULTS: The results indicated that SO2 treatment maintained the postharvest quality of table grapes. The sulfite content in rachises and berries, but not the sulfate content, increased in response to SO2 treatment. SO2 caused high activities of sulfite reductase, O-acetylserine (thiol)-lyase, and γ-glutamylcysteine synthetase, thereby increasing the contents of cysteine, hydrogen sulfide, and glutathione in the rachises and berries. The expression of VvSURTL, VvATPS1, VvATPS2, and VvAPR3 decreased in response to SO2 treatment; however, the transcript levels of VvSiR1 and VvOASTL exhibited the opposite tendency. CONCLUSION: These findings indicated that the sulfite converted from SO2 participated in sulfur metabolism and maintained the postharvest quality of table grapes by modulating the contents of metabolites, activities of enzymes, and expression of genes related to sulfur metabolism. © 2021 Society of Chemical Industry.


Sulfites/metabolism , Sulfur Dioxide/pharmacology , Sulfur/metabolism , Vitis/metabolism , Fruit/chemistry , Fruit/drug effects , Fruit/metabolism , Sulfites/analysis , Sulfur/analysis , Sulfur Dioxide/metabolism , Vitis/chemistry , Vitis/drug effects
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