In silico identification of genes involved in selenium metabolism: evidence for a third selenium utilization trait

<p>Abstract</p> <p>Background</p> <p>Selenium (Se) is a trace element that occurs in proteins in the form of selenocysteine (Sec) and in tRNAs in the form of selenouridine (SeU). Selenophosphate synthetase (SelD) is required for both utilization traits. However, previous research also revealed SelDs in two organisms lacking Sec and SeU, suggesting a possible additional use of Se that is dependent on SelD.</p> <p>Results</p> <p>In this study, we conducted comparative genomics and phylogenetic analyses to characterize genes involved in Se utilization. Candidate genes identified included SelA/SelB and YbbB that define Sec and SeU pathways, respectively, and NADH oxidoreductase that is predicted to generate a SelD substrate. In addition, among 227 organisms containing SelD, 10 prokaryotes were identified that lacked SelA/SelB and YbbB. Investigation of <it>selD </it>neighboring genes in these organisms revealed a SirA-like protein and two hypothetical proteins HP1 and HP2 that were strongly linked to a novel Se utilization. With these new signature proteins, 32 bacteria and archaea were found that utilized these proteins, likely as part of the new Se utilization trait. Metabolic labeling of one organism containing an orphan SelD, <it>Enterococcus faecalis</it>, with ⁷⁵Se revealed a protein containing labile Se species that could be released by treatment with reducing agents, suggesting non-Sec utilization of Se in this organism.</p> <p>Conclusion</p> <p>These studies suggest the occurrence of a third Se utilization trait in bacteria and archaea.</p

UGA codon position-dependent incorporation of selenocysteine into mammalian selenoproteins

It is thought that the SelenoCysteine Insertion Sequence (SECIS) element and UGA codon are sufficient for selenocysteine (Sec) insertion. However, we found that UGA supported Sec insertion only at its natural position or in its close proximity in mammalian thioredoxin reductase 1 (TR1). In contrast, Sec could be inserted at any tested position in mammalian TR3. Replacement of the 3′-UTR of TR3 with the corresponding segment of a Euplotes crassus TR restricted Sec insertion into the C-terminal region, whereas the 3′-UTR of TR3 conferred unrestricted Sec insertion into E. crassus TR, in which Sec insertion is normally limited to the C-terminal region. Exchanges of 3′-UTRs between mammalian TR1 and E. crassus TR had no effect, as both proteins restricted Sec insertion. We further found that these effects could be explained by the use of selenoprotein-specific SECIS elements. Examination of Sec insertion into other selenoproteins was consistent with this model. The data indicate that mammals evolved the ability to limit Sec insertion into natural positions within selenoproteins, but do so in a selenoprotein-specific manner, and that this process is controlled by the SECIS element in the 3′-UTR

Adjustments, extinction, and remains of selenocysteine incorporation machinery in the nematode lineage

This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date. After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International).Selenocysteine (Sec) is encoded by an UGA codon with the help of a SECIS element present in selenoprotein mRNAs. SECIS-binding protein (SBP2/SCBP-2) mediates Sec insertion, but the roles of its domains and the impact of its deficiency on Sec insertion are not fully understood. We used Caenorhabditis elegans to examine SBP2 function since it possesses a single selenoprotein, thioredoxin reductase-1 (TRXR-1). All SBP2 described so far have an RNA-binding domain (RBD) and a Secincorporation domain (SID). Surprisingly, C. elegans SBP2 lacks SID and consists only of an RBD. An sbp2 deletion mutant strain ablated Sec incorporation demonstrating SBP2 essentiality for Sec incorporation. Further in silico analyses of nematode genomes revealed conservation of SBP2 lacking SID and maintenance of Sec incorporation linked to TRXR-1. Remarkably, parasitic plant nematodes lost the ability to incorporate Sec, but retained SecP43, a gene associated with Sec incorporation. Interestingly, both selenophosphate synthetase (SPS) genes are absent in plant parasitic nematodes, while only Cys-containing SPS2 is present in Sec-incorporating nematodes. Our results indicate that C. elegans and the nematode lineage provide key insights into Sec incorporation and the evolution of Sec utilization trait, selenoproteomes, selenoproteins, and Sec residues. Finally, our study provides evidence of noncanonical translation initiation in C. elegans, not previously known for this well-established animal model.This work was supported by Universidad de la República, Uruguay (Grant Number 418 to G.S., PhD fellowship to L.O.); Asociación Española de Cooperación Internacional (C/7646/07 to A.M.-V. and G.S.; A/016083/08 to A.M.-V. and G.S.); Asociación Universitaria Iberoamericana de Posgrado and Agencia Nacional de Innovación e Investigación (BE_POS_2009_183 and BE_POS_2010_2160 to L.O.), and was partially funded by FOCEM (MERCOSUR Structural Convergence Fund), [COF 03/11].Peer Reviewe

Regulation of Selenocysteine Content of Human Selenoprotein P by Dietary Selenium and Insertion of Cysteine in Place of Selenocysteine

Selenoproteins are a unique group of proteins that contain selenium in the form of selenocysteine (Sec) co-translationally inserted in response to a UGA codon with the help of cis- and trans-acting factors. Mammalian selenoproteins contain single Sec residues, with the exception of selenoprotein P (SelP) that has 7–15 Sec residues depending on species. Assessing an individual’s selenium status is important under various pathological conditions, which requires a reliable selenium biomarker. Due to a key role in organismal selenium homeostasis, high Sec content, regulation by dietary selenium, and availability of robust assays in human plasma, SelP has emerged as a major biomarker of selenium status. Here, we found that Cys is present in various Sec positions in human SelP. Treatment of cells expressing SelP with thiophosphate, an analog of the selenium donor for Sec synthesis, led to a nearly complete replacement of Sec with Cys, whereas supplementation of cells with selenium supported Sec insertion. SelP isolated directly from human plasma had up to 8% Cys inserted in place of Sec, depending on the Sec position. These findings suggest that a change in selenium status may be reflected in both SelP concentration and its Sec content, and that availability of the SelP-derived selenium for selenoprotein synthesis may be overestimated under conditions of low selenium status due to replacement of Sec with Cys

Differentiation of Transbaikal Territory by Tick-Borne Viral Encephalitis Incidence

Objective of the study was to differentiate the Administrative Districts of theTransbaikalTerritory at the time of Tick-borne viral encephalitis (TBVE) incidence recession by epidemiological risk groups and to characterize them by volume of specific and nonspecific preventive measures.Materials and methods. Retrospective analysis of TBVE epidemiological situation is based on the statistical reporting data “Information on infectious and parasitic diseases” in 2009–2019 and other materials of the Rospotrebnadzor Administration in the Transbaikal Territory. The clustering of areas with various levels of epidemiological risk was conducted by calculation of 95 % confidential interval for long-term annual average of TBVE cases in municipal units of the Territory over a decade and assessment of appurtenance of the deviating values to the aggregate under study.Results and discussion. Twenty four out of 32 districts of the Transbaikal Territory are endemic for TBVE. These areas are divided into five groups: with very high epidemiological risk (2 districts), high (5), medium (8), and low (8) risk respectively, as well as the administrative center of the constituent entity which by the whole complex of indicators (disease manifestation, population density, factors of targeted TBVE decrease, social-and-living and economical conditions) cannot be considered together with the rest of municipalities. Each group of the districts was characterized by the number of cases and TBVE incidence rates, medical aid seeking by persons who suffered from tick bites, vaccination volumes, seroprevention, areas of acaricide treatments. Recommendations are presented for the essential complex and scope of measures to prevent TBVE in the groups of administrative districts that differ by the level of epidemiological risk

Analyses of Fruit Flies That Do Not Express Selenoproteins or Express the Mouse Selenoprotein, Methionine Sulfoxide Reductase B1, Reveal a Role of Selenoproteins in Stress Resistance

Selenoproteins are essential in vertebrates because of their crucial role in cellular redox homeostasis, but some invertebrates that lack selenoproteins have recently been identified. Genetic disruption of selenoprotein biosynthesis had no effect on lifespan and oxidative stress resistance of Drosophila melanogaster. In the current study, fruit flies with knock-out of the selenocysteine-specific elongation factor were metabolically labeled with 75Se; they did not incorporate selenium into proteins and had the same lifespan on a chemically defined diet with or without selenium supplementation. These flies were, however, more susceptible to starvation than controls, and this effect could be ascribed to the function of selenoprotein K. We further expressed mouse methionine sulfoxide reductase B1 (MsrB1), a selenoenzyme that catalyzes the reduction of oxidized methionine residues and has protein repair function, in the whole body or the nervous system of fruit flies. This exogenous selenoprotein could only be expressed when the Drosophila selenocysteine insertion sequence element was used, whereas the corresponding mouse element did not support selenoprotein synthesis. Ectopic expression of MsrB1 in the nervous system led to an increase in the resistance against oxidative stress and starvation, but did not affect lifespan and reproduction, whereas ubiquitous MsrB1 expression had no effect. Dietary selenium did not influence lifespan of MsrB1-expressing flies. Thus, in contrast to vertebrates, fruit flies preserve only three selenoproteins, which are not essential and play a role only under certain stress conditions, thereby limiting the use of the micronutrient selenium by these organisms

Clinical-Epidemiological Peculiarities of the Tick-Borne Borrelioses Registered in the Trans-Baikal Territory

Complex analysis of the data on epidemiology and clinical picture of the tick-borne borrelioses in the territory of the Trans-Baikal Region over the last decade since 2003 to 2012 has demonstrated that there is a distinct upward tendency as concerns its morbidity rates. Spotted have been the potentially hazardous, as regards the infection, areas. Highest incidence rates are registered between May-July among adult men, and erythema form of the disease prevails. Based on the results of molecular-genetic investigation of Ixodidae ticks, for the first time ever in the territory of the Dul’durginsk Region identified has been circulation of Borrelia garinii , and B. afzelii , pathogenic for humans bacterial species of Borrelia genus

Epidemiological Situation on Tick-Borne Viral Encephalitis in the Russian Federation in 2011–2021 and Short-Term Forecast of its Development

The aim of the work was to analyze the epidemiological situation on tick-borne viral encephalitis in the endemic territories of the Russian Federation in 2021 in comparison with the dynamics over 2011–2020 and its shortterm forecast for 2022. In Russia, 48 constituent entities belonging to seven federal districts are endemic for tick-borne viral encephalitis (TBVE). There is a statistically significant downward trend in the incidence of TBVE in the Siberian Federal District (which is characterized by the maximum incidence rate in the country), the Volga and Far Eastern Federal Districts. In the Ural Federal District (the second in terms of TBVE incidence), the decline in the incidence has stalled since 2021. The average long-term incidence of TBVE in the constituent entities of the Russian Federation varies from sporadic (Republic of Crimea)  up to 12.5 per 100 thousand of the population (0/0000) (Krasnoyarsk Territory). In 2021, TBVE cases were detected in 42 endemic regions and in one non-endemic region – Stavropol Territory. At the same time, 1015 cases of TBVE were in the country (0.69 0/0000). In all Federal Districts, the incidence of TBVE is below the long-term average values. Using the Quantum GIS program, the incidence of TBVE in 917 administrative territories of the country has been ranked and grouped according to the level of epidemiological risk. This made it possible to establish that 65 % of the territories form a zone of low epidemiological risk. High and very high epidemiological risk is observed in 13% of the analyzed districts. The structure of TBVE clinical manifestations in 2021 was dominated by febrile (59.7 %) and meningeal (24.3 %) forms. 14 lethal outcomes were reported. In 2021, 2 889 515 people were vaccinated (including 1 433 850 children), of which 14 fell ill. Specific immunoglobulin was used to prevent the overt development of infection in 100 704 individuals, which accounts for 22.6 % of the persons affected by tick bites (30.6 % among children). Acaricidic treatments were carried out on an operational area of 233 125 hectares of territories of socially significant objects. The scope of all TBVE prevention measures in 2021 increased as compared to 2020. In 2022, a decline in the incidence of TBVE in endemic Federal Districts and in the country on the whole is forecasted to (0,64±0,192) 0/0000

Reduced Utilization of Selenium by Naked Mole Rats Due to a Specific Defect in GPx1 Expression

Naked mole rat (MR) Heterocephalus glaber is a rodent model of delayed aging because of its unusually long life span (\u3e28 years). It is also not known to develop cancer. In the current work, tissue imaging by x-ray fluorescence microscopy and direct analyses of trace elements revealed low levels of selenium in the MR liver and kidney, whereas MR and mouse brains had similar selenium levels. This effect was not explained by uniform selenium deficiency because methionine sulfoxide reductase activities were similar in mice and MR. However, glutathione peroxidase activity was an order of magnitude lower inMRliver and kidney than in mouse tissues. In addition, metabolic labeling of MR cells with 75Se revealed a loss of the abundant glutathione peroxidase 1 (GPx1) band, whereas other selenoproteins were preserved. To characterize theMRselenoproteome, we sequenced its liver transcriptome. Gene reconstruction revealed standard selenoprotein sequences except for GPx1, which had an early stop codon, and SelP, which had low selenocysteine content. When expressed inHEK293cells,MRGPx1waspresent in low levels,and its expression could be rescued neither by removing the early stop codon nor by replacing its SECIS element. In addition, GPx1 mRNAwas present in lower levels inMRliver than in mouse liver. To determine if GPx1 deficiency could account for the reduced selenium content, we analyzed GPx1 knock-out mice and found reduced selenium levels in their livers and kidneys. Thus, MR is characterized by the reduced utilization of selenium due to a specific defect in GPx1 expression

Diversity of Protein and mRNA Forms of Mammalian Methionine Sulfoxide Reductase B1 Due to Intronization and Protein Processing

Background: Methionine sulfoxide reductases (Msrs) are repair enzymes that protect proteins from oxidative stress by catalyzing stereospecific reduction of oxidized methionine residues. MsrB1 is a selenocysteine-containing cytosolic/nuclear Msr with high expression in liver and kidney. Principal Findings: Here, we identified differences in MsrB1 gene structure among mammals. Human MsrB1 gene consists of four, whereas the corresponding mouse gene of five exons, due to occurrence of an additional intron that flanks the stop signal and covers a large part of the 3′-UTR. This intron evolved in a subset of rodents through intronization of exonic sequences, whereas the human gene structure represents the ancestral form. In mice, both splice forms were detected in liver, kidney, brain and heart with the five-exon form being the major form. We found that both mRNA forms were translated and supported efficient selenocysteine insertion into MsrB1. In addition, MsrB1 occurs in two protein forms that migrate as 14 and 5 kDa proteins. We found that each mRNA splice form generated both protein forms. The abundance of the 5 kDa form was not influenced by protease inhibitors, replacement of selenocysteine in the active site or mutation of amino acids in the cleavage site. However, mutation of cysteines that coordinate a structural zinc decreased the levels of 5 and 14 kDa forms, suggesting importance of protein structure for biosynthesis and/stability of these forms. Conclusions: This study characterized unexpected diversity of protein and mRNA forms of mammalian selenoprotein MsrB1