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1.
Cyril Moccand Svetlana Boycheva Pedro Surriabre Marina Tambasco-Studart Maja Raschke Markus Kaufmann Teresa B. Fitzpatrick 《The Journal of biological chemistry》2014,289(12):8203-8216
Vitamin B6 is an indispensable compound for survival, well known as a cofactor for numerous central metabolic enzymes and more recently for playing a role in several stress responses, particularly in association with oxidative stress. Regulatory aspects for the use of the vitamin in these roles are not known. Here we show that certain plants carry a pseudoenzyme (PDX1.2), which is involved in regulating vitamin B6 biosynthesis de novo under stress conditions. Specifically, we demonstrate that Arabidopsis PDX1.2 enhances the activity of its catalytic paralogs by forming a heterododecameric complex. PDX1.2 is strongly induced by heat as well as singlet oxygen stress, concomitant with an enhancement of vitamin B6 production. Analysis of pdx1.2 knockdown lines demonstrates that boosting vitamin B6 content is dependent on PDX1.2, revealing that this pseudoenzyme acts as a positive regulator of vitamin B6 biosynthesis during such stress conditions in plants. 相似文献
2.
Nathalie Mangel Jared B. Fudge Kuan‐Te Li Ting‐Ying Wu Takayuki Tohge Alisdair R. Fernie Boris Szurek Teresa B. Fitzpatrick Wilhelm Gruissem Herv Vanderschuren 《The Plant journal : for cell and molecular biology》2019,99(6):1047-1065
Vitamin B6 (pyridoxine) is vital for key metabolic reactions and reported to have antioxidant properties in planta. Therefore, enhancement of vitamin B6 content has been hypothesized to be a route to improve resistance to biotic and abiotic stresses. Most of the current studies on vitamin B6 in plants are on eudicot species, with monocots remaining largely unexplored. In this study, we investigated vitamin B6 biosynthesis in rice, with a view to examining the feasibility and impact of enhancing vitamin B6 levels. Constitutive expression in rice of two Arabidopsis thaliana genes from the vitamin B6 biosynthesis de novo pathway, AtPDX1.1 and AtPDX2, resulted in a considerable increase in vitamin B6 in leaves (up to 28.3‐fold) and roots (up to 12‐fold), with minimal impact on general growth. Rice lines accumulating high levels of vitamin B6 did not display enhanced tolerance to abiotic stress (salt) or biotic stress (resistance to Xanthomonas oryzae infection). While a significant increase in vitamin B6 content could also be achieved in rice seeds (up to 3.1‐fold), the increase was largely due to its accumulation in seed coat and embryo tissues, with little enhancement observed in the endosperm. However, seed yield was affected in some vitamin B6‐enhanced lines. Notably, expression of the transgenes did not affect the expression of the endogenous rice PDX genes. Intriguingly, despite transgene expression in leaves and seeds, the corresponding proteins were only detectable in leaves and could not be observed in seeds, possibly pointing to a mode of regulation in this organ. 相似文献
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4.
Functional analysis of PDX2 from Arabidopsis, a glutaminase involved in vitamin B6 biosynthesis
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Vitamin B6 is an essential metabolite in all organisms, being required as a cofactor for a wide variety of biochemical reactions. De novo biosynthesis of the vitamin occurs in microorganisms and plants, but animals must obtain it from their diet. Two distinct and mutually exclusive de novo pathways have been identified to date, namely deoxyxylulose 5-phosphate dependent, which is restricted to a subset of eubacteria, and deoxyxylulose 5-phosphate independent, present in archaea, fungi, plants, protista, and most eubacteria. In these organisms, pyridoxal 5'-phosphate (PLP) formation is catalyzed by a single glutamine amidotransferase (PLP synthase) composed of a glutaminase domain, PDX2, and a synthase domain, PDX1. Despite plants being an important source of vitamin B6, very little is known about its biosynthesis. Here, we provide information for Arabidopsis thaliana. The functionality of PDX2 is demonstrated, using both in vitro and in vivo analyses. The expression pattern of PDX2 is assessed at both the RNA and protein level, providing insight into the spatial and temporal pattern of vitamin B6 biosynthesis. We then provide a detailed biochemical analysis of the plant PLP synthase complex. While the active sites of PDX1 and PDX2 are remote from each other, coordination of catalysis is much more pronounced with the plant proteins than its bacterial counterpart, Bacillus subtilis. Based on a model of the PDX1/PDX2 complex, mutation of a single residue uncouples enzyme coordination and in turn provides tangible evidence for the existence of the recently proposed ammonia tunnel through the core of PDX1. 相似文献
5.
Maite Colinas Marion Eisenhut Takayuki Tohge Marta Pesquera Alisdair R. Fernie Andreas P.M. Weber Teresa B. Fitzpatrick 《The Plant cell》2016,28(2):439-453
Vitamin B6 comprises a family of compounds that is essential for all organisms, most notable among which is the cofactor pyridoxal 5′-phosphate (PLP). Other forms of vitamin B6 include pyridoxamine 5′-phosphate (PMP), pyridoxine 5′-phosphate (PNP), and the corresponding nonphosphorylated derivatives. While plants can biosynthesize PLP de novo, they also have salvage pathways that serve to interconvert the different vitamers. The selective contribution of these various pathways to cellular vitamin B6 homeostasis in plants is not fully understood. Although biosynthesis de novo has been extensively characterized, the salvage pathways have received comparatively little attention in plants. Here, we show that the PMP/PNP oxidase PDX3 is essential for balancing B6 vitamer levels in Arabidopsis thaliana. In the absence of PDX3, growth and development are impaired and the metabolite profile is altered. Surprisingly, RNA sequencing reveals strong induction of stress-related genes in pdx3, particularly those associated with biotic stress that coincides with an increase in salicylic acid levels. Intriguingly, exogenous ammonium rescues the growth and developmental phenotype in line with a severe reduction in nitrate reductase activity that may be due to the overaccumulation of PMP in pdx3. Our analyses demonstrate an important link between vitamin B6 homeostasis and nitrogen metabolism. 相似文献
6.
Main conclusion
PDX1.2 is expressed in the basal part of the globular-stage embryo, and plays critical roles in development, hypocotyl elongation, and stress response.Abstract
The Arabidopsis thaliana PDX1.2 protein belongs to a small family of three members. While PDX1.1 and PDX1.3 have been extensively described and are well established to function in vitamin B6 biosynthesis, the biological role of PDX1.2 still remains elusive. Here, we show that PDX1.2 is expressed early in embryo development, and that heat shock treatment causes a strong up-regulation of the gene. Using a combined genetic approach of T-DNA insertion lines and expression of artificial micro RNAs, we can show that PDX1.2 is critically required for embryo development, and for normal hypocotyl elongation. Plants with reduced PDX1.2 expression also display reduced primary root growth after heat shock treatments. The work overall provides a set of important new findings that give greater insights into the developmental role of PDX1.2 in plants. 相似文献7.
Michel Havaux Brigitte Ksas Agnieszka Szewczyk Dominique Rumeau Fabrice Franck Stefano Caffarri Christian Triantaphylidès 《BMC plant biology》2009,9(1):130
Background
Vitamin B6 is a collective term for a group of six interconvertible compounds: pyridoxine, pyridoxal, pyridoxamine and their phosphorylated derivatives. Vitamin B6 plays essential roles as a cofactor in a range of biochemical reactions. In addition, vitamin B6 is able to quench reactive oxygen species in vitro, and exogenously applied vitamin B6 protects plant cells against cell death induced by singlet oxygen (1O2). These results raise the important question as to whether plants employ vitamin B6 as an antioxidant to protect themselves against reactive oxygen species. 相似文献8.
Yoshikazu Izumi Kuninori Sato Yoshiki Tani Koichi Ogata 《Bioscience, biotechnology, and biochemistry》2013,77(11):2683-2684
Vitamin B6 is synthesized by green Cytisus scoparius callus and green Phellodendron amurense callus cultured on Linsmaier and Skoog Agar-medium with 10?5m of ±-naphthaleneacetic acid (NAA) and 10?6 m of 6-benzyladenine (BA). Even when thiamine and inositol were omitted from this medium, the growth and vitamin B6 content of Cytisus scoparius callus did not change. Vitamin B6 contents of clones of the calluses varied and were unstable during long-term subculture. Clonal selection was repeated to obtain stable strains with high vitamin B6 content, and the vitamin B6 content of one strain of green Cytisus scoparius callus became 4-times higher than that of the green leaves. 相似文献
9.
Naomichi Nishio Kenji Sakai Koji Fujii Tadashi Kamikubo 《Bioscience, biotechnology, and biochemistry》2013,77(3):553-559
The accumulation of vitamin B6 by Pichia guilliermondii Wickerham NK–2 strain grown on hydrocarbon was investigated. Ammonium acetate was more effective than other nitrogen sources tested. Satisfactory utilization by the yeast strain was observed in n-alkanes of C10–C18, and n-pentadecane was the best for vitamin B6 production. Vitamin B6 was excreted in the cultural broth mainly in the form of pyridoxal, The maximal vitamin B6 production was approximately 25 mg per liter of the culture broth. 相似文献
10.
Plants of two varieties of winter wheat—Besostoya 1 and Etoile de Choisie— were raised in the field. The same number of selected plants from each variety were frozen once in a refrigerator at a temperature of ?15°C for 24 hrs. After freezing, the plants were gradually thawed and returned to the field. Early in the spring, test and control plants were treated with one of following diluted solutions: vitamin B1, vitamin B6, indolyl-3-acetic acid and adenine (the four solutions at a concentration of 0·01% and 5% urea solution in water) by immersion for 15 hrs. at a temperature of 18–20°C. The plants were measured in the spring and in the summer, the yield being determined after ripening. It was found that all substances used helped to diminish the deleterious consequences of overwintering and to raise the yield of the plants. Vitamin B6 exerted the strongest influence. Vitamin B6 treated plants produced higher or slightly lower yields than those of the non-frozen control. Plants treated with vitamin B1 gave yields close to, but in most cases lower than those of the non-frozen control. The yields of plants treated with solutions of the remaining substances were appreciably lower than those of the non-frozen controls. Plants of the more winter-hardy variety Besostoya 1 recovered relatively more easily under the effect of the substances used than the plants of the less winter-hardy variety Etoile de Choise. 相似文献
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12.
The seasonal and spatial distribution of vitamin B12 was investigated in Eastern Mediterranean (Saronicos Gulf, Aegean Sea). Vitamin B12 concentration had an annual range 0.12–7.93 ng. 1−1 and an annual mean 1.88 ng. 1−1. The statistical analysis of data indicated significant seasonal and regional variations of vitamin B12 in this area and an inverse relationship of this vitamin with chl a. The results showed that the Eastern Mediterranean Sea contains less vitamin B12 than the Western Mediterranean Sea. 相似文献
13.
Nicolas Szydlowski Lukas Bürkle Lucille Pourcel Michael Moulin Jürgen Stolz Teresa B. Fitzpatrick 《The Plant journal : for cell and molecular biology》2013,75(1):40-52
Vitamin B6 is a cofactor for more than 140 essential enzymatic reactions and was recently proposed as a potent antioxidant, playing a role in the photoprotection of plants. De novo biosynthesis of the vitamin has been described relatively recently and is derived from simple sugar precursors as well as glutamine. In addition, the vitamin can be taken up from exogenous sources in a broad range of organisms, including plants. However, specific transporters have been identified only in yeast. Here we assess the ability of the family of Arabidopsis purine permeases (PUPs) to transport vitamin B6. Several members of the family complement the growth phenotype of a Saccharomyces cerevisiae mutant strain impaired in both de novo biosynthesis of vitamin B6 as well as its uptake. The strongest activity was observed with PUP1 and was confirmed by direct measurement of uptake in yeast as well as in planta, defining PUP1 as a high affinity transporter for pyridoxine. At the tissue level the protein is localised to hydathodes and here we use confocal microscopy to illustrate that at the cellular level it is targeted to the plasma membrane. Interestingly, we observe alterations in pyridoxine recycling from the guttation sap upon overexpression of PUP1 and in a pup1 mutant, consistent with the role of the protein in retrieval of pyridoxine. Furthermore, combining the pup1 mutant with a vitamin B6 de novo biosynthesis mutant (pdx1.3) corroborates that PUP1 is involved in the uptake of the vitamin. 相似文献
14.
Trayambak Basak Gaurav Garg Nitin Bhardwaj Vinay Singh Tanwar Sandeep Seth Ganesan Karthikeyan 《Biomarkers》2016,21(5):436-440
Coronary artery disease (CAD) has been increasing alarmingly in India. We had earlier shown that vitamin B12 deficiency is associated with CAD in Indian population. However, only about a quarter of the total vitamin B12 is internalised in the cells by the proteins transcobalamin II. Vitamin B12-bound transcobalamin II (holotranscobalamin, holoTC) is thus referred to as biologically active B12. In this study, we ascertained the levels of holoTC in 501 CAD cases and 1253 healthy controls and for the first time show that holoTC levels are significantly lower (p?=?2.57E-4) in CAD (26.81?pmol/l) cases as compared to controls (29.97?pmol/l). 相似文献
15.
Vinay Singh Tanwar Mandeep P. Chand Jitender Kumar Gaurav Garg Sandeep Seth Ganesan Karthikeyan Shantanu Sengupta 《Gene》2013
Vitamin B12 is an essential micronutrient synthesized by microorganisms. Mammals including humans have evolved ways for transport and absorption of this vitamin. Deficiency of vitamin B12 (either due to low intake or polymorphism in genes involved in absorption and intracellular transport of this vitamin) has been associated with various complex diseases. Genome-wide association studies have recently identified several common single nucleotide polymorphisms (SNPs) in fucosyl transferase 2 gene (FUT2) to be associated with levels of vitamin B12—the strongest association was with a non-synonymous SNP rs602662 in this gene. In the present study, we attempted to replicate the association of this SNP (rs602662) in an Indian population since a significant proportion has been reported to have low levels of vitamin B12 in this population. A total of 1146 individuals were genotyped for this SNP using a single base extension method and association with levels of vitamin B12 was assessed in these individuals. Regression analysis was performed to analyze the association considering various confounding factors like for age, sex, diet, hypertension, diabetes mellitus and coronary artery disease status. We found that the SNP rs602662 was significantly associated with the levels of vitamin B12 (p value < 0.0001). We also found that individuals adhering to a vegetarian diet with GG (homozygous major genotype) have significantly lower levels of vitamin B12 in these individuals. Thus, our study reveals that vegetarian diet along with polymorphism in the FUT2 gene may contribute significantly to the high prevalence of vitamin B12 deficiency in India. 相似文献
16.
Vitamin B6 is an essential nutrient in the human diet. It can act as a co-enzyme for numerous metabolic enzymes and has recently been shown to be a potent antioxidant. Plants and microorganisms have the ability to make the compound. Yet, studies of vitamin B6 biosynthesis have been mainly restricted to Escherichia coli, where the vitamin is synthesized from 1-deoxy-d -xylulose 5-phosphate and 4-phosphohydroxy-l-threonine. Recently, a novel pathway for its synthesis has been discovered, involving two genes (PDX1 and PDX2) neither of which is homologous to any of those participating in the E. coli pathway. In Bacillus subtilis, YaaD and YaaE represent the PDX1 and PDX2 homolog, respectively. The two proteins form a complex that functions as a glutamine amidotransferase, with YaaE as the glutaminase domain and YaaD as the acceptor and pyridoxal 5'-phosphate (PLP) synthesis domain. In this report we corroborate a recent report on the identification of the substrates of YaaD and provide unequivocal proof of the identity of the reaction product. We show that both the glutaminase and synthase reactions are dependent on the respective protein partner. The synthase reaction can also utilize an external ammonium source but, in contrast to other glutamine amidotransferases, is dependent on YaaE under certain conditions. Furthermore, we report on the detailed characterization of the inhibition of the glutaminase domain, and thus PLP synthesis, by the glutamine analog acivicin. Employing pull-out assays and native-PAGE, we provide evidence for the dissociation of the bi-enzyme complex under these conditions. The results are discussed in light of the nature of the interaction of the two components of the enzyme complex. 相似文献
17.
Analysis of the Arabidopsis rsr4-1/pdx1-3 mutant reveals the critical function of the PDX1 protein family in metabolism, development, and vitamin B6 biosynthesis
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Wagner S Bernhardt A Leuendorf JE Drewke C Lytovchenko A Mujahed N Gurgui C Frommer WB Leistner E Fernie AR Hellmann H 《The Plant cell》2006,18(7):1722-1735
Vitamin B6 represents a highly important group of compounds ubiquitous in all living organisms. It has been demonstrated to alleviate oxidative stress and in its phosphorylated form participates as a cofactor in >100 biochemical reactions. By means of a genetic approach, we have identified a novel mutant, rsr4-1 (for reduced sugar response), with aberrant root and leaf growth that requires supplementation of vitamin B6 for normal development. Cloning of the mutated gene revealed that rsr4-1 carries a point mutation in a member of the PDX1/SOR1/SNZ (for Pyridoxine biosynthesis protein 1/Singlet oxygen resistant 1/Snooze) family that leads to reduced vitamin B6 content. Consequently, metabolism is broadly altered, mainly affecting amino acid, raffinose, and shikimate contents and trichloroacetic acid cycle constituents. Yeast two-hybrid and pull-down analyses showed that Arabidopsis thaliana PDX1 proteins can form oligomers. Interestingly, the mutant form of PDX1 has severely reduced capability to oligomerize, potentially suggesting that oligomerization is important for function. In summary, our results demonstrate the critical function of the PDX1 protein family for metabolism, whole-plant development, and vitamin B6 biosynthesis in higher plants. 相似文献
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《Insect Biochemistry》1984,14(2):175-179
Vitamin B12 concentrations were determined by radioassay in the housefly, five species of termites, and 17 other phylogenetically diverse insect species. Vitamin B12 was not detected in the housefly Musca domestica, which apparently cannot interconvert propionate and succinate. In contrast, the termite Zootermopsis angusticollis readily interconverts succinate and methylmalonate, and contains high amounts of vitamin B12 (940 pg/mg dry tissue), as do four other species of termites. Experiments involving selective elimination of either gut tract protozoa or bacteria in Coptotermes formosanus indicate that intestinal bacteria are the major source of vitamin B12 in this termite. The other insect species examined have undetectable to moderate amounts of vitamin B12. 相似文献
20.
Lipid profile of the spinal cord myelin was studied in normal and vitamin B12 deficient chicks. The significant findings were a reduction in the total galactolipids and an increase in the total phospholipids of myelin in vitamin B12 deficiency. The altered molar ratios of these lipids suggest a relative immaturity of the myelin in this condition. These changes may initiate the degenerative changes in the central nervous system in vitamin B12 deficiency.Vitamin B12 is essential for normal functioning of both the hemopoietic and the nervous system. Sub-acute combined degeneration of the spinal cord is seen both in association with pernicious anemia and in megaloblastic anemia of dietary origin (1,2). Though many biochemical postulates (3, 4) have been advanced to explain the neurological changes, the role of vitamin B12 in maintaining the integrity of myelin is still obscure. So far only in two animal species the monkey (5) and the chick (6) has myelin degeneration been reported in vitamin B12 deficiency. However, in neither of these reports, have changes in the composition of myelin been described. The results of a study in chicks wherein the effects of vitamin B12 on the lipid profile of myelin were investigated are reported here. 相似文献