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result(s) for
"monosaccharide"
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Engineering of yeast hexose transporters to transport d-xylose without inhibition by d-glucose
by
Bruder, Stefan
,
Farwick, Alexander
,
Schadeweg, Virginia
in
Amino acids
,
asparagine
,
Biological Sciences
2014
All known d -xylose transporters are competitively inhibited by d -glucose, which is one of the major reasons hampering simultaneous fermentation of d -glucose and d -xylose, two primary sugars present in lignocellulosic biomass. We have set up a yeast growth-based screening system for mutant d -xylose transporters that are insensitive to the presence of d -glucose. All of the identified variants had a mutation at either a conserved asparagine residue in transmembrane helix 8 or a threonine residue in transmembrane helix 5. According to a homology model of the yeast hexose transporter Gal2 deduced from the crystal structure of the d -xylose transporter XylE from Escherichia coli , both residues are found in the same region of the protein and are positioned slightly to the extracellular side of the central sugar-binding pocket. Therefore, it is likely that alterations sterically prevent d -glucose but not d -xylose from entering the pocket. In contrast, changing amino acids that are supposed to directly interact with the C6 hydroxymethyl group of d -glucose negatively affected transport of both d -glucose and d -xylose. Determination of kinetic properties of the mutant transporters revealed that Gal2-N376F had the highest affinity for d -xylose, along with a moderate transport velocity, and had completely lost the ability to transport hexoses. These transporter versions should prove valuable for glucose–xylose cofermentation in lignocellulosic hydrolysates by Saccharomyces cerevisiae and other biotechnologically relevant organisms. Moreover, our data contribute to the mechanistic understanding of sugar transport because the decisive role of the conserved asparagine residue for determining sugar specificity has not been recognized before.
Journal Article
Hxt1, a monosaccharide transporter and sensor required for virulence of the maize pathogen Ustilago maydis
by
Norbert Sauer
,
Martin Münsterkötter
,
Miroslav Vranes
in
Amino Acid Substitution
,
avirulent strains
,
biotrophic development
2015
The smut Ustilago maydis, a ubiquitous pest of corn, is highly adapted to its host to parasitize on its organic carbon sources. We have identified a hexose transporter, Hxt1, as important for fungal development during both the saprophytic and the pathogenic stage of the fungus.
Hxt1 was characterized as a high-affinity transporter for glucose, fructose, and mannose; ∆hxt1 strains show significantly reduced growth on these substrates, setting Hxt1 as the main hexose transporter during saprophytic growth.
After plant infection, ∆hxt1 strains show decreased symptom development. However, expression of a Hxt1 protein with a mutation leading to constitutively active signaling in the yeast glucose sensors Snf3p and Rgt2p results in completely apathogenic strains. Fungal development is stalled immediately after plant penetration, implying a dual function of Hxt1 as transporter and sensor.
As glucose sensors are only known for yeasts, ‘transceptor’ as Hxt1 may constitute a general mechanism for sensing of glucose in fungi. In U. maydis, Hxt1 links a nutrient-dependent environmental signal to the developmental program during pathogenic development.
Journal Article
Molecular Identification and Physiological Characterization of a Novel Monosaccharide Transporter from Arabidopsis Involved in Vacuolar Sugar Transport
by
Trentmann, Oliver
,
Tjaden, Joachim
,
Schmidt, Ulrike
in
Amino Acid Sequence
,
Arabidopsis
,
Arabidopsis - cytology
2006
The tonoplast monosaccharide transporter (TMT) family comprises three isoforms in Arabidopsis thaliana, and TMT-green fluorescent protein fusion proteins are targeted to the vacuolar membrane. TMT promoter-β-glucuronidase plants revealed that the TONOPLAST MONOSACCHARIDE TRANSPORTER1 (TMT1) and TMT2 genes exhibit a tissue- and cell type-specific expression pattern, whereas TMT3 is only weakly expressed. TMT1 and TMT2 expression is induced by drought, salt, and cold treatments and by sugar. During cold adaptation, tmt knockout lines accumulated less glucose and fructose compared with wild-type plants, whereas no differences were observed for sucrose. Cold adaptation of wild-type plants substantially promoted glucose uptake into isolated leaf mesophyll vacuoles. Glucose uptake into isolated vacuoles was inhibited by NH₄⁺, fructose, and phlorizin, indicating that transport is energy-dependent and that both glucose and fructose were taken up by the same carrier. Glucose import into vacuoles from two cold-induced tmt1 knockout lines or from triple knockout plants was substantially lower than into corresponding wild-type vacuoles. Monosaccharide feeding into leaf discs revealed the strongest response to sugar in tmt1 knockout lines compared with wild-type plants, suggesting that TMT1 is required for cytosolic glucose homeostasis. Our results indicate that TMT1 is involved in vacuolar monosaccharide transport and plays a major role during stress responses.
Journal Article
Sugar transporters in Fabaceae, featuring SUT MST and SWEET families of the model plant Medicago truncatula and the agricultural crop Pisum sativum
2019
Sugar transporters play a crucial role for plant productivity, as they coordinate sugar fluxes from source leaf towards sink organs (seed, fruit, root) and regulate the supply of carbon resources towards the microorganisms of the rhizosphere (bacteria and fungi). Thus, sugar fluxes mediated by SUT (sucrose transporters), MST (monosaccharide transporters) and SWEET (sugar will eventually be exported transporters) families are key determinants of crop yield and shape the microbial communities living in the soil. In this work, we performed a systematic search for sugar transporters in Fabaceae genomes, focusing on model and agronomical plants. Here, we update the inventory of sugar transporter families mining the latest version of the Medicago truncatula genome and identify for the first time SUT MST and SWEET families of the agricultural crop Pisum sativum. The sugar transporter families of these Fabaceae species comprise respectively 7 MtSUT 7 PsSUT, 72 MtMST 59 PsMST and 26 MtSWEET 22 PsSWEET. Our comprehensive phylogenetic analysis sets a milestone for the scientific community, as we propose a new and simple nomenclature to correctly name SUT MST and SWEET families. Then, we searched for transcriptomic data available for our gene repertoire. We show that several clusters of homologous genes are co-expressed in different organs, suggesting that orthologous sugar transporters may have a conserved function. We focused our analysis on gene candidates that may be involved in remobilizing resources during flowering, grain filling and in allocating carbon towards roots colonized by arbuscular mycorrhizal fungi and Rhizobia. Our findings open new perspectives for agroecological applications in legume crops, as for instance improving the yield and quality of seed productions and promoting the use of symbiotic microorganisms.
Journal Article
A Versatile Monosaccharide Transporter That Operates in the Arbuscular Mycorrhizal Fungus Glomus sp Is Crucial for the Symbiotic Relationship with Plants
by
Wippel, Kathrin
,
Requena, Natalia
,
Helber, Nicole
in
Base Sequence
,
Biological Transport
,
Carbon
2011
For more than 400 million years, plants have maintained a mutualistic symbiosis with arbuscular mycorrhizal (AM) fungi. This evolutionary success can be traced to the role of these fungi in providing plants with mineral nutrients, particularly phosphate. In return, photosynthates are given to the fungus, which support its obligate biotrophic lifestyle. Although the mechanisms involved in phosphate transfer have been extensively studied, less is known about the reciprocal transfer of carbon. Here, we present the high-affinity Monosaccharide Transporter2 (MST2) from Glomus sp with a broad substrate spectrum that functions at several symbiotic root locations. Plant cell wall sugars can efficiently outcompete the Glc uptake capacity of MST2, suggesting they can serve as alternative carbon sources. MST2 expression closely correlates with that of the mycorrhiza-specific Phosphate Transported (PT4). Furthermore, reduction of MST2 expression using host-induced gene silencing resulted in impaired mycorrhiza formation, malformed arbuscules, and reduced PT4 expression. These findings highlight the symbiotic role of MST2 and support the hypothesis that the exchange of carbon for phosphate is tightly linked. Unexpectedly, we found that the external mycelium of AM fungi is able to take up sugars in a proton-dependent manner. These results imply that the sugar uptake system operating in this symbiosis is more complex than previously anticipated.
Journal Article
Plant glucose transporter structure and function
2020
The carbohydrate D-glucose is the main source of energy in living organisms. In contrast to animals, as well as most fungi, bacteria, and archaea, plants are capable to synthesize a surplus of sugars characterizing them as autothrophic organisms. Thus, plants are de facto the source of all food on earth, either directly or indirectly via feed to livestock. Glucose is stored as polymeric glucan, in animals as glycogen and in plants as starch. Despite serving a general source for metabolic energy and energy storage, glucose is the main building block for cellulose synthesis and represents the metabolic starting point of carboxylate- and amino acid synthesis. Finally yet importantly, glucose functions as signalling molecule conveying the plant metabolic status for adjustment of growth, development, and survival. Therefore, cell-to-cell and long-distance transport of photoassimilates/sugars throughout the plant body require the fine-tuned activity of sugar transporters facilitating the transport across membranes. The functional plant counterparts of the animal sodium/glucose transporters (SGLTs) are represented by the proton-coupled sugar transport proteins (STPs) of the plant monosaccharide transporter(-like) family (MST). In the framework of this special issue on “Glucose Transporters in Health and Disease,” this review gives an overview of the function and structure of plant STPs in comparison to the respective knowledge obtained with the animal Na+-coupled glucose transporters (SGLTs).
Journal Article
Structural basis for transport and inhibition of the human glucose-6-phosphate transporter G6PT
2025
The human glucose-6-phosphate transporter (G6PT) moves glucose-6-phosphate (G6P) into the lumen of endoplasmic reticulum, playing a vital role in glucose homeostasis. Dysregulation of G6PT causes glycogen storage disease 1b. Despite its functional importance, the structure, G6P recognition, and inhibition mechanism of G6PT remain unclear. Here, we report the cryo-EM structures of human G6PT in apo, G6P-bound, and the specific inhibitor chlorogenic acid (CHA)-bound forms, elucidating the structural basis for G6PT transport and inhibition. The G6P pocket comprises subsite A for phosphate and subsite B for glucose. The CHA occupies the G6P site and locks G6PT in a partly-occluded state. Functional assays demonstrate that G6PT activity is enhanced by co-expression of glucose-6-phosphatase (G6PC), but G6PT does not form a complex with G6PC. Together, this study provides a solid foundation for understanding the structure‒function relationships and pathology of G6PT and sheds light on the future development of potential therapeutics targeting G6PT.
G6PT plays a vital role in glucose homeostasis by transporting glucose-6-phosphate to the lumen of ER. Here, authors present the cryo-EM structures of human G6PT in distinct forms, revealing the structural basis for transport and inhibition of G6PT.
Journal Article
Regulation of sugar transporter activity for antibacterial defense in Arabidopsis
by
Takano, Yoshiaka
,
Yamada, Kohji
,
Nakagami, Hirofumi
in
Amino Acid Sequence
,
Arabidopsis
,
Arabidopsis - metabolism
2016
Microbial pathogens strategically acquire metabolites from their hosts during infection. Here we show that the host can intervene to prevent such metabolite loss to pathogens. Phosphorylation-dependent regulation of sugar transport protein 13 (STP13) is required for antibacterial defense in the plant Arabidopsis thaliana. STP13 physically associates with the flagellin receptor flagellin-sensitive 2 (FLS2) and its co-receptor BRASSINOSTEROID INSENSITIVE 1-associated receptor kinase 1 (BAK1). BAK1 phosphorylates STP13 at threonine 485, which enhances its monosaccharide uptake activity to compete with bacteria for extracellular sugars. Limiting the availability of extracellular sugar deprives bacteria of an energy source and restricts virulence factor delivery. Our results reveal that control of sugar uptake, managed by regulation of a host sugar transporter, is a defense strategy deployed against microbial infection. Competition for sugar thus shapes host-pathogen interactions.
Journal Article
CRISPR/Cas9-mediated mutation of OsSWEET14 in rice cv. Zhonghua11 confers resistance to Xanthomonas oryzae pv. oryzae without yield penalty
2020
Background
Bacterial blight of rice, caused by
Xanthomonas oryzae
pv.
oryzae
(
Xoo
), is a devastating rice disease in Southeast Asia and West Africa.
OsSWEET14
, encoding a sugar transporter, is known to be a major susceptible gene of bacterial blight targeted by four different transcription activator-like (TAL) effectors from either Asian or African
Xoo
strains. However, the
OsSWEET14
single knockout or promoter mutants in the Kitaake background are moderately resistant or even susceptible to African
Xoo
strains. Therefore, in this study, we knocked out
OsSWEET14
in rice cv. Zhonghua 11 background for disease assessment.
Results
In this study, CRISPR/Cas9 was utilized to disrupt the function of
OsSWEET14
by modifying its corresponding coding region in the genome of rice cv. Zhonghua 11 (
CR-S14
). In total, we obtained nine different
OsSWEET14
-mutant alleles. Besides conferring broad-spectrum resistance to Asian
Xoo
strains, tested mutant alleles also showed strong resistance to African
Xoo
strain AXO1947. Moreover, the expression of
OsSWEET14
was detected in vascular tissues, including the stem, leaf sheath, leaf blade and root. The disruption of
OsSWEET14
led to increased plant height without a reduction in yield.
Conclusions
Disruption of
OsSWEET14
in the Zhonghua 11 background is able to confer strong resistance to African
Xoo
strain AXO1947 and Asian
Xoo
strain PXO86.
CR-S14
has normal reproductive growth and enhanced plant height under normal growth conditions. These results imply that
CR-S14
may serve as a better tester line than
sweet14
single-knockout mutant in the Kitaake background for the diagnostic kit for rice blight resistance. The genetic background and increased plant height need to be taken into consideration when utilizing
OsSWEET14
for resistant rice breeding.
Journal Article
NRT/PTR transporters are essential for translocation of glucosinolate defence compounds to seeds
2012
Two high-affinity proton-dependent transporters of glucosinolates have been identified in
Arabidopsis
and termed GTR1 and GTR2; these transporters are essential for transporting glucosinolates to seeds, offering a means to control the allocation of defence compounds in a tissue-specific manner, which may have agricultural biotechnology implications.
Engineering more nutritional crops
Glucosinolates are important plant defence compounds. They are synthesized in various tissues and then translocated to the seeds, where they accumulate. In this study, Barbara Halkier and colleagues examine the molecular basis of this long-distance transport process. They identify two high-affinity, proton-dependent glucosinolate-specific transporters in
Arabidopsis
, termed GTR1 and GTR2. These transporters control the loading of glucosinolates from the apoplast into the phloem. The authors' specific and complete elimination of glucosinolates from
Arabidopsis
seeds, combined with the compounds' retention in vegetative tissues, establishes transport engineering as a potential approach for eliminating anti-nutritional natural products in high-value crops.
In plants, transport processes are important for the reallocation of defence compounds to protect tissues of high value
1
, as demonstrated in the plant model
Arabidopsis
, in which the major defence compounds, glucosinolates
2
, are translocated to seeds on maturation
3
. The molecular basis for long-distance transport of glucosinolates and other defence compounds, however, remains unknown. Here we identify and characterize two members of the nitrate/peptide transporter family, GTR1 and GTR2, as high-affinity, proton-dependent glucosinolate-specific transporters. The
gtr1 gtr2
double mutant did not accumulate glucosinolates in seeds and had more than tenfold over-accumulation in source tissues such as leaves and silique walls, indicating that both plasma membrane-localized transporters are essential for long-distance transport of glucosinolates. We propose that GTR1 and GTR2 control the loading of glucosinolates from the apoplasm into the phloem. Identification of the glucosinolate transporters has agricultural potential as a means to control allocation of defence compounds in a tissue-specific manner.
Journal Article