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Summary: In plants under drought stress, sugar content in roots increases, which is important for drought resistance. However, the molecular mechanisms for controlling the sugar content in roots during response to drought remain elusive. Here, we found that the MdDOF3‐MdHT1.2 module‐mediated glucose influx into the root is essential for drought resistance in apple (Malus × domestica). Drought induced glucose uptake from the rhizosphere and up‐regulated the transcription of hexose transporter MdHT1.2. Compared with the wild‐type plants, overexpression of MdHT1.2 promoted glucose uptake from the rhizosphere, thereby facilitating sugar accumulation in root and enhancing drought resistance, whereas silenced plants showed the opposite phenotype. Furthermore, ATAC‐seq, RNA‐seq and biochemical analysis demonstrated that MdDOF3 directly bound to the promoter of MdHT1.2 and was strongly up‐regulated under drought. Overexpression of MdDOF3 in roots improved MdHT1.2‐mediated glucose transport capacity and enhanced plant resistance to drought, but MdDOF3‐RNAihr apple plants showed the opposite phenotype. Moreover, overexpression of MdDOF3 in roots did not attenuate drought sensitivity in MdHT1.2‐RNAi plants, which was correlated with a lower glucose uptake capacity and glucose content in root. Collectively, our findings deciphered the molecular mechanism through which glucose uptake from the rhizosphere is mediated by MdDOF3‐MdHT1.2, which acts to modulate sugar content in root and promote drought resistance. [ABSTRACT FROM AUTHOR]

Pollen cells require large amounts of sugars from the anther to support their development, which is critical for plant sexual reproduction and crop yield. Sugars Will Eventually be Exported Transporters (SWEETs) have been shown to play an important role in the apoplasmic unloading of sugars from anther tissues into symplasmically isolated developing pollen cells and thereby affect the sugar supply for pollen development. However, among the 17 CsSWEET genes identified in the cucumber (Cucumis sativus L.) genome, the CsSWEET gene involved in this process has not been identified. Here, a member of the SWEET gene family, CsSWEET5a, was identified and characterized. The quantitative real-time PCR and β-glucuronidase expression analysis revealed that CsSWEET5a is highly expressed in the anthers and pollen cells of male cucumber flowers from the microsporocyte stage (stage 9) to the mature pollen stage (stage 12). Its subcellular localization indicated that the CsSWEET5a protein is localized to the plasma membrane. The heterologous expression assays in yeast demonstrated that CsSWEET5a encodes a hexose transporter that can complement both glucose and fructose transport deficiencies. CsSWEET5a can significantly rescue the pollen viability and fertility of atsweet8 mutant Arabidopsis plants. The possible role of CsSWEET5a in supplying hexose to developing pollen cells via the apoplast is also discussed. [ABSTRACT FROM AUTHOR]

【Objective】Hexose transporter (HXT) is a kind of monosaccharide transporter, which can transport mannose, fructose, glucose and other hexose substances across the membrane, and participate in the development of pollen and the growth of the whole plant, and then determine the internal quality of fruit. Bioinformatics and expression characteristics of HXT gene in pitaya were analyzed in order to provide a theoretical basis for regulating the fruit quality of pitaya.【Method】A hexose transporter gene was obtained by gene cloning. Bioinformatics analysis was conducted by using NCBI, BioEdit, DNAMAN and other software or databases. The soluble sugar contents were determined by HPLC at fruit maturity stage. The relative expression characteristics of HTX gene in different lines of pitaya and different parts of afruit were analyzed by fluorescence quantitative PCR.【Result】A HXT gene named HuHXTwas cloned from pitaya fruit by PCR amplification. The total length of the gene was 1 230 bp, encoding 410 amino acids. The protein instability coefficient was 35.59, with 32 Ser, 15 Thr and 15 Tyr. There were sucrose, glucose and fructose in pitaya, and glucose and fructose were the main components. The expression of HuHXT was the highest in No. 97 line, followed by Zhanhong No. 2 and No. 128, while No. 15 and No. 16 had the lowest expression level. The expression level of HuHXT was not significantly different in different parts of a fruit. The expression in parts near the pericarp was higher other parts of a fruit.【Conclusion】HuHXT is a member of the Sugar-tr gene family. HuHXT protein is a stable protein with strong hydrophobicity. As a membrane protein, its secondary structure is mainly α-helix and random curl. There is negative correlation between the relative expression of HuHXT and the content of glucose, that is, the higher the HuHXT expression level, the lower the glucose content.

Despite intense elimination efforts, human malaria, caused by the infection of five Plasmodium species, remains the deadliest parasitic disease in the world. Even worse, with the emergence and spreading of the first‐line drug‐resistant Plasmodium parasites, therapeutic interventions based on novel plasmodial drug targets are more necessary than ever. Given that the blood‐stage parasites primarily rely on glycolysis for their energy supply, blocking glucose uptake, the rate‐limiting step of ATP generation, was considered a promising approach to kill these parasites. To achieve this goal, characterization of the plasmodial hexose transporter and development of selective inhibitors have been pursued for decades. Here, we review the identification and characterization of the Plasmodium falciparum hexose transporter (PfHT1) and summarize current advances in its inhibitor development. [ABSTRACT FROM AUTHOR]

Sugars, which are critical osmotic compounds and signalling molecules in plants, and Sugars Will Eventually be Exported Transporters (SWEETs), which constitute a novel family of sugar transporters, play central roles in plant responses to multiple abiotic stresses. In the present study, a member of the SWEET gene family from cucumber (Cucumis sativus L.), CsSWEET2, was identified and characterized. Histochemical analysis of β-glucuronidase expression in transgenic Arabidopsis plants showed that CsSWEET2 is highly expressed in the leaves; subcellular localization indicated that CsSWEET2 proteins are localized in the plasma membrane and endoplasmic reticulum. Heterologous expression assays in yeast demonstrated that CsSWEET2 encodes an energy-independent hexose/H+ uniporter that can complement both glucose and fructose transport deficiencies. Compared with wild-type Arabidopsis plants, transgenic Arabidopsis plants overexpressing CsSWEET2 had much lower relative electrolyte leakage levels and were much more resistant to cold stress. Sugar content analysis showed that glucose and fructose levels in the transgenic Arabidopsis plants were significantly higher than those in the wild-type plants. Taken together, our results suggest that, by mediating sugar metabolism and compartmentation, CsSWEET2 plays a vital role in improving plant cold tolerance. [ABSTRACT FROM AUTHOR]

During anthesis, there is an increased demand for carbohydrates due to pollen maturation and nectary secretion that warrants a systematic phloem unloading strategy for sugar partitioning. Sugar transporters are key components of the apoplasmic phloem unloading strategy and control the sugar flux needed for plant development. Currently, the phloem unloading strategy during anthesis has not been explored in cucumber, and the question of which sugar transporters are active during flower anthesis is poorly understood. In this study, a study utilizing the phloem-mobile symplasmic tracer carboxyfluorescein (CF) suggested that the phloem unloading was symplasmically isolated in the receptacle and nectary of cucumber flowers at anthesis. We also identified a hexose transporter that is highly expressed in cucumber flower, Sugar Will Eventually be Exported Transporter 7a (SWEET7a). CsSWEET7a was mainly expressed in receptacle and nectary tissues in both male and female flowers, where its expression level increased rapidly right before anthesis. At anthesis, the CsSWEET7a protein was specifically localized to the phloem region of the receptacle and nectary, indicating that CsSWEET7a may function in the apoplasmic phloem unloading during flower anthesis. Although cucumber mainly transports raffinose family oligosaccharides (RFOs) in the phloem, sucrose, glucose, and fructose are the major sugars in the flower receptacle and the nectary as well as in nectar at anthesis. In addition, the transcript levels of genes encoding soluble sugar hydrolases (α-galactosidase, sucrose synthase, cytoplasmic invertase, and cell wall invertase) were correlated with that of CsSWEET7a. These results indicated that CsSWEET7a may be involved in sugar partitioning as an exporter in the phloem of the receptacle and nectary to supply carbohydrates for flower anthesis and nectar secretion in cucumber.

During anthesis, there is an increased demand for carbohydrates due to pollen maturation and nectary secretion that warrants a systematic phloem unloading strategy for sugar partitioning. Sugar transporters are key components of the apoplasmic phloem unloading strategy and control the sugar flux needed for plant development. Currently, the phloem unloading strategy during anthesis has not been explored in cucumber, and the question of which sugar transporters are active during flower anthesis is poorly understood. In this study, a study utilizing the phloem-mobile symplasmic tracer carboxyfluorescein (CF) suggested that the phloem unloading was symplasmically isolated in the receptacle and nectary of cucumber flowers at anthesis. We also identified a hexose transporter that is highly expressed in cucumber flower, Sugar Will Eventually be Exported Transporter 7a (SWEET7a). CsSWEET7a was mainly expressed in receptacle and nectary tissues in both male and female flowers, where its expression level increased rapidly right before anthesis. At anthesis, the CsSWEET7a protein was specifically localized to the phloem region of the receptacle and nectary, indicating that CsSWEET7a may function in the apoplasmic phloem unloading during flower anthesis. Although cucumber mainly transports raffinose family oligosaccharides (RFOs) in the phloem, sucrose, glucose, and fructose are the major sugars in the flower receptacle and the nectary as well as in nectar at anthesis. In addition, the transcript levels of genes encoding soluble sugar hydrolases (α -galactosidase , sucrose synthase , cytoplasmic invertase , and cell wall invertase) were correlated with that of CsSWEET7a. These results indicated that CsSWEET7a may be involved in sugar partitioning as an exporter in the phloem of the receptacle and nectary to supply carbohydrates for flower anthesis and nectar secretion in cucumber. [ABSTRACT FROM AUTHOR]

Aims: In this study, we attempted to increase the productivity of Candida glycerinogenes yeast for ethanol production from non‐detoxified sugarcane bagasse hydrolysates (NDSBH) by identifying the hexose transporter in this yeast that makes a high contribution to glucose consumption, and by adding additional copies of this transporter and enhancing its membrane localisation stability (MLS). Methods and Results: Based on the knockout and overexpression of key hexose transporter genes and the characterisation of their promoter properties, we found that Cghxt4 and Cghxt6 play major roles in the early and late stages of fermentation, respectively, with Cghxt4 contributing most to glucose consumption. Next, subcellular localisation analysis revealed that a common mutation of two ubiquitination sites (K9 and K538) in Cghxt4 improved its MLS. Finally, we overexpressed this Cghxt4 mutant (Cghxt4.2A) using a strong promoter, PCgGAP, which resulted in a significant increase in the ethanol productivity of C. glycerinogenes in the NDSBH medium. Specifically, the recombinant strain showed 18 and 25% higher ethanol productivity than the control in two kinds of YP‐NDSBH medium (YP‐NDSBH1G160 and YP‐NDSBH2G160), respectively. Conclusions: The hexose transporter mutant Cghxt4.2A (Cghxt4K9A,K538A) with multiple copies and high MLS was able to significantly increase the ethanol productivity of C. glycerinogenes in NDSBH. Significance and Impact of the Study: Our results provide a promising strategy for constructing efficient strains for ethanol production. [ABSTRACT FROM AUTHOR]

Efficient hexose transporters are essential for the development of industrial yeast strains with high fermentation performance. We previously identified a hexose transporter, CgHxt4, with excellent sugar uptake performance at ultra-high glucose concentrations (200 g/L) in the high sugar fermenting yeast C. glycerinogenes. To understand the working mechanism of this transporter, we constructed 87 mutants and examined their glucose uptake performance. The results revealed that five residues (N321, N322, F325, G426, and P427) are essential for the efficient glucose transport of CgHxt4. Subsequently, we focused our analysis on the roles of N321 and P427. Specifically, N321 and P427 are likely to play a role in glucose coordination and conformational flexibility, respectively. Our results help to expand the application potential of this transporter and provide insights into the working mechanism of yeast hexose transporter. Key points: • Five residues, transmembrane segments 7 and 10, were found to be essential for CgHxt4. • N321 and P427 are likely to play a role in glucose coordination and conformational flexibility, respectively. • Chimeric CgHxt5.4TM7 significantly enhanced the performance of CgHxt5. [ABSTRACT FROM AUTHOR]

Early fruit development is critical for determining crop yield. Cell wall invertase (CWIN) and sugar transporters both play important roles in carbon allocation and plant development. However, there is little information about the relationship between CWIN and those functionally related sugar transporters during fruit development. By using transgenic tomato with an elevated CWIN activity, we investigated how an increase in CWIN activity may regulate the expression of sugar transporter genes during fruit development. Our analyses indicate that CWIN activity may be under tight regulation by multiple regulators, including two invertase inhibitors (INVINHs) and one defective CWIN (deCWIN) in tomato ovaries prior to anthesis. Among the sugar transporters, expression of SlSWEET12c for sucrose efflux and SlHT2 for hexose uptake was enhanced by the elevated CWIN activity at 10 and 15 days after anthesis of tomato fruit development, respectively. The findings show that some specific sugars will eventually be exported transporters (SWEETs) and hexose transporters (HTs) respond to elevate CWIN activity probably to promote rapid fruit expansion when sucrose efflux from phloem and hexose uptake by parenchyma cell are in high demand. The analyses provide new leads for improving crop yield by manipulating CWIN-responsive sugar transporters, together with CWIN itself, to enhance fruit development and sugar accumulation.

Yeast and cancer cells are metabolically similar as they use fermentation of glucose as a primary means of generating energy. Reliance on glucose fermentation makes both of these cell types highly sensitive to the toxic glucose analog, 2-deoxyglucose. Here we review the cellular and metabolic pathways that play a role in 2-deoxyglucose sensitivity and discuss how the modifications to these pathways result in acquisition of 2-deoxyglucose resistance. Insights gained from genetic and proteomic studies in yeast provide new ideas for the design of combinatorial therapies for cancer treatment. [ABSTRACT FROM AUTHOR]

Artemisinin-resistant malaria parasites have emerged and have been spreading, posing a significant public health challenge. Antimalarial drugs with novel mechanisms of action are therefore urgently needed. In this report, we exploit a "selective starvation" strategy by inhibiting Plasmodium falciparum hexose transporter 1 (PfHT1), the sole hexose transporter in P. falciparum, over human glucose transporter 1 (hGLUT1), providing an alternative approach to fight against multidrug-resistant malaria parasites. The crystal structure of hGLUT3, which shares 80% sequence similarity with hGLUT1, was resolved in complex with C3361, a moderate PfHT1-specific inhibitor, at 2.3-Å resolution. Structural comparison between the present hGLUT3-C3361 and our previously reported PfHT1-C3361 confirmed the unique inhibitor binding-induced pocket in PfHT1. We then designed small molecules to simultaneously block the orthosteric and allosteric pockets of PfHT1. Through extensive structure-activity relationship studies, the TH-PF series was identified to selectively inhibit PfHT1 over hGLUT1 and potent against multiple strains of the blood-stage P. falciparum. Our findings shed light on the next-generation chemotherapeutics with a paradigm-shifting structure-based design strategy to simultaneously target the orthosteric and allosteric sites of a transporter. [ABSTRACT FROM AUTHOR]

Salt is one of the main stresses that limit plant growth, especially at the seedling stage, reducing crop production and severely impacting food security. However, the relationship between salt stress and sugar content regulated by sugar transporters remains unknown. Here, we investigated the salt tolerance of transgenic tomato seedlings ectopically expressing MdHT2.2 , which is a fructose and glucose/H+ symporter located on the plasma membrane in apple. Although the contents of fructose, glucose and sucrose in the leaves of seedlings ectopically expressing MdHT2.2 obviously increased compared with those of WT seedlings, the transgenic seedlings were significantly less tolerance to salt stress. Under salt stress, the SlSOS1/2 and SlNHX1 genes were highly expressed, and the accumulation of Na+ was lower in the transgenic seedlings than in WT, however, ROS accumulated to a greater degree in the former, and the ROS-scavenging-related enzyme activities and AsA content were lower in the transgenic seedlings than WT. Taken together, these results indicated that the relatively low salt tolerance of the MdHT2.2 transgenic seedlings was related with the accumulation of ROS, which was caused by reduced ROS-scavenging ability. Our results offer proof that changes in sugar content caused by sugar transporters are related to salt tolerance, and provide new insight into the regulation of sugar content, quality improvement and stress tolerance. Image 1 • MdHT2.2 transgenic seedlings accumulated more sugar than WT. • MdHT2.2 transgenic seedlings showed lower salt resistance but had less Na+ accumulation. • MdHT2.2 transgenic seedlings accumulated more ROS accompanied with lower ROS scavenging ability. • The sugar content change caused by MdHT2.2 ectopic expression influenced the ROS scavenging system sensing salt stress. [ABSTRACT FROM AUTHOR]

Eukaryotic genomes contain numerous homologous repeat sequences including redundant genes with divergent homology that can be potential recombination targets. Recombination between divergent sequences is rare but poses a substantial threat to genome stability. The hexose transporter (HXT) gene family shares high sequence similarities at both protein and DNA levels, and some members are placed close together in tandem arrays. In this study, we show that spontaneous interstitial deletions occur at significantly high rates in HXT gene clusters, resulting in chimeric HXT sequences that contain a single junction point. We also observed that DNA double-strand breaks created between HXT genes produce primarily interstitial deletions, whereas internal cleavage of the HXT gene resulted in gene conversions as well as deletion products. Interestingly, interstitial deletions were less constrained by sequence divergence than gene conversion. Moreover, recombination-defective mutations differentially affected the survival frequency. Mutations that impair single-strand annealing (SSA) pathway greatly reduced the survival frequency by 10–1,000-fold, whereas disruption of Rad51-dependent homologous recombination exhibited only modest reduction. Our results indicate that recombination in the tandemly repeated HXT genes occurs primarily via SSA pathway. [ABSTRACT FROM AUTHOR]

Target of rapamycin (TOR) kinases form two distinct complexes, TORC1 and TORC2, which are evolutionarily conserved among eukaryotes. These complexes control intracellular biochemical processes in response to changes in extracellular nutrient conditions. Previous studies using the fission yeast, Schizosaccharomyces pombe, showed that the TORC2 signaling pathway, which is essential for cell proliferation under glucose-limited conditions, ensures cell-surface localization of a high-affinity hexose transporter, Ght5, by downregulating its endocytosis. The TORC2 signaling pathway retains Ght5 on the cell surface, depending on the presence of nitrogen sources in medium. Ght5 is transported to vacuoles upon nitrogen starvation. In this review, we discuss the molecular mechanisms underlying this regulation to cope with nutritional stress, a response which may be conserved from yeasts to mammals.

Summary: Sugar transporters are necessary to transfer hexose from cell wall spaces into parenchyma cells to boost hexose accumulation to high concentrations in fruit. Here, we have identified an apple hexose transporter (HTs), MdHT2.2, located in the plasma membrane, which is highly expressed in mature fruit. In a yeast system, the MdHT2.2 protein exhibited high 14C‐fructose and 14C‐glucose transport activity. In transgenic tomato heterologously expressing MdHT2.2, the levels of both fructose and glucose increased significantly in mature fruit, with sugar being unloaded via the apoplastic pathway, but the level of sucrose decreased significantly. Analysis of enzyme activity and the expression of genes related to sugar metabolism and transport revealed greatly up‐regulated expression of SlLIN5, a key gene encoding cell wall invertase (CWINV), as well as increased CWINV activity in tomatoes transformed with MdHT2.2. Moreover, the levels of fructose, glucose and sucrose recovered nearly to those of the wild type in the sllin5‐edited mutant of the MdHT2.2‐expressing lines. However, the overexpression of MdHT2.2 decreased hexose levels and increased sucrose levels in mature leaves and young fruit, suggesting that the response pathway for the apoplastic hexose signal differs among tomato tissues. The present study identifies a new HTs in apple that is able to take up fructose and glucose into cells and confirms that the apoplastic hexose levels regulated by HT controls CWINV activity to alter carbohydrate partitioning and sugar content. [ABSTRACT FROM AUTHOR]

Hexose transporters play many important roles in plant development. However, the role of hexose transporter in secondary cell wall growth has not been reported before. Here, we report that the hexose transporter gene CsHT3 is mainly expressed in cells with secondary cell walls in cucumber. Spatiotemporal expression analysis revealed that the transcript of CsHT3 mainly accumulates in the stem, petiole, tendril, and peduncle, all of which contain high cellulose levels. Immunolocalization results show that CsHT3 is localized at the sclereids in young peduncles, shifts to the phloem fiber cells during peduncle development, and then shifts again to the companion cells when the development of secondary cell walls is almost completed. Carboxyfluoresce unloading experiment indicated that carbohydrate unloading in the phloem follows an apoplastic pathway. Overexpression of CsHT3 in cucumber plant can improve the cellulose content and cell wall thickness of phloem fiber cells in the peduncle. The expression of cellulose synthase genes were increased in the CsHT3 overexpression plants. These results indicated that CsHT3 may play an important role in cellulose synthesis through promoting the expression of cellulose synthase genes. • Phloem unloading in the cucumber peduncles following an apoplastic pathway. • The localization of CsHT3 transfer to different cell types of phloem during peduncle development. • First identified the function of a hexose transporter in peduncle. • Overexpression of CsHT3 improved the cellulose content and cell wall thickness of phloem fiber cells in cucumber peduncle. [ABSTRACT FROM AUTHOR]

During high gravity fermentation, a set of hexose transporters in yeasts plays an important role in efficient sugar transport. However, hexose transporters have been studied mainly in the Saccharomyces cerevisiae model and at low or moderate sugar concentrations. The hexose transporters are still poorly understood in the industrial glycerol producer Candida glycerinogenes, which assimilates sugar efficiently at high glucose concentration. To explore these hexose transporters, 14 candidates were identified using a hidden Markov model and characterized. Five of these functioned as hexose transporters when expressed in S. cerevisiae. In particular, CgHxt4 showed the highest efficiency of glucose transport at elevated glucose concentration among a group of transporters including Hxt1 and Hxt7 from S. cerevisiae. qRT-PCR in C. glycerinogenes revealed that transcription of CgHXT4 was induced by high glucose concentrations while fluorescence localization analysis indicated that CgHxt4 remained relatively stable on the membrane under these conditions. In addition, site-directed mutagenesis revealed that the asparagine 329 from CgHxt4, located in the YYX(T/P) conserved motif of hexose transporters, promoted an increased glucose transport. Overexpressing CgHXT4 in S. cerevisiae enhanced glucose consumption and ethanol production more effectively at high glucose concentrations than ScHXT1, the most significant native transporter from S. cerevisiae. These results indicate that CgHxt4 plays an important role in the fermentation process as a hexose transporter with strong transport activity and efficient expression regulation at high glucose concentrations. [ABSTRACT FROM AUTHOR]

Microbial fitness is easy to measure in the laboratory, but difficult to measure in the field. Laboratory fitness assays make use of controlled conditions and genetically modified organisms, neither of which are available in the field. Among other applications, fitness assays can help researchers detect adaptation to different habitats or locations. We designed a competitive fitness assay to detect adaptation of Saccharomyces paradoxus isolates to the habitat they were isolated from (oak or larch leaf litter). The assay accurately measures relative fitness by tracking genotype frequency changes in the field using digital droplet PCR ( DDPCR). We expected locally adapted S. paradoxus strains to increase in frequency over time when growing on the leaf litter type from which they were isolated. The DDPCR assay successfully detected fitness differences among S. paradoxus strains, but did not find a tendency for strains to be adapted to the habitat they were isolated from. Instead, we found that the natural alleles of the hexose transport gene we used to distinguish S. paradoxus strains had significant effects on fitness. The origin of a strain also affected its fitness: strains isolated from oak litter were generally fitter than strains from larch litter. Our results suggest that dispersal limitation and genetic drift shape S. paradoxus populations in the forest more than local selection does, although further research is needed to confirm this. Tracking genotype frequency changes using DDPCR is a practical and accurate microbial fitness assay for natural environments. [ABSTRACT FROM AUTHOR]

Understanding the mechanism of photosynthate transfer at symbiotic interface by fungal monosaccharide transporter is of substantial importance. The carbohydrate uptake at the apoplast by the fungus is facilitated by PiHXT5 hexose transporter in root endophytic fungus Piriformospora indica. The putative PiHXT5 belongs to MFS superfamily with twelve predicted transmembrane helices. It possesses sugar transporter PFAM motif (PF0083) and MFS superfamily domain (PS50850). It contains the signature tags related to glucose transporter GLUT1 of human erythrocyte. PiHXT5 is regulated in response to mutualism as well as glucose concentration. We have functionally characterized PiHXT5 by complementation of hxt-null mutant of Saccharomyces cerevisiae EBY.VW4000. It is involved in transport of multiple sugars ranging from D-glucose, D-fructose, D-xylose, D-mannose, D-galactose with decreasing affinity. The uncoupling experiments indicate that it functions as H+/glucose co-transporter. Further, pH dependence analysis suggests that it functions maximum between pH 5 to 6. The expression of PiHXT5 is dependent on glucose concentration and was found to be expressed at low glucose levels (1 mM) which indicate its role as a high affinity glucose transporter. Our study on this sugar transporter will help in better understanding of carbon metabolism and flow in this agro-friendly fungus.

Candida glabrata is an emerging human fungal pathogen that has efficacious nutrient sensing and responsiveness ability. It can be seen through its ability to thrive in diverse range of nutrient limited-human anatomical sites. Therefore, nutrient sensing particularly glucose sensing is thought to be crucial in contributing to the development and fitness of the pathogen. This study aimed to elucidate the role of SNF3 (Sucrose Non Fermenting 3) as a glucose sensor and its possible role in contributing to the fitness and survivability of C. glabrata in glucose-limited environment. The SNF3 knockout strain was constructed and subjected to different glucose concentrations to evaluate its growth, biofilm formation, amphotericin B susceptibility, ex vivo survivability and effects on the transcriptional profiling of the sugar receptor repressor (SRR) pathway-related genes. The SNF3Δ strain showed a retarded growth in low glucose environments (0.01% and 0.1%) in both fermentation and respiration-preferred conditions but grew well in high glucose concentration environments (1% and 2%). It was also found to be more susceptible to amphotericin B in low glucose environment (0.1%) and macrophage engulfment but showed no difference in the biofilm formation capability. The deletion of SNF3 also resulted in the down-regulation of about half of hexose transporters genes (4 out of 9). Overall, the deletion of SNF3 causes significant reduction in the ability of C. glabrata to sense limited surrounding glucose and consequently disrupts its competency to transport and perform the uptake of this critical nutrient. This study highlighted the role of SNF3 as a high affinity glucose sensor and its role in aiding the survivability of C. glabrata particularly in glucose limited environment.

This trial was conducted to investigate the effects of feeding system and age on morphological development (including villus height, crypt depth, and ratio of villus height to crypt depth) and expression of hexose transporters (i.e., SGLT1, GLUT2 and GLUT5) in different segments of the small intestine of goat kids. A total of 40 Liuyang black goat kids were randomly slaughtered at 0, 7, 28, 42, 56 and 70 days of age. There were two feeding systems [i.e., supplemental feeding (S) and grazing (G)] from 28 to 70 days of age. Duodenal, jejunal and ileal tissues were collected to measure the villus height and crypt depth. Mucosa samples were taken to quantify mRNA or protein expression of sodium-dependent glucose-linked transporter-1 (SGLT1), facilitated glucose/fructose transporter 2 (GLUT2), and facilitated glucose transporter 5 (GLUT5). Villus height was not affected ( P > 0.05) by feeding system, and crypt depth was only affected ( P < 0.001) by feeding system within the jejunum. Moreover, villus height increased ( P = 0.006) with increasing age in the duodenum, whereas it decreased as age increased in the jejunum ( P = 0.040) and ileum ( P < 0.001). Additionally, the jejunal and ileal crypt depth linearly increased ( P < 0.001) with increasing age in both S and G groups. The feeding system had a significant effect ( P < 0.05) on mRNA expression of SGLT1, GLUT2 and GLUT5 in the intestine, except for GLUT2 expression within the ileum. The mRNA and protein expression of SGLT1 and the mRNA expression of GLUT2 and GLUT5 declined ( P < 0.001) with increasing age in the jejunum and ileum. The present results indicate that supplemental feeding at an early age is beneficial to activate expression of SGLT1, GLUT2 and GLUT5 transporters in goats for an improved ability to absorb hexose. [ABSTRACT FROM AUTHOR]

Understanding the mechanism of photosynthate transfer at symbiotic interface by fungal monosaccharide transporter is of substantial importance. The carbohydrate uptake at the apoplast by the fungus is facilitated by PiHXT5 hexose transporter in root endophytic fungus Piriformospora indica. The putative PiHXT5 belongs to MFS superfamily with 12 predicted transmembrane helices. It possess sugar transporter PFAM motif (PF0083) and MFS superfamily domain (PS50850). It contains the signature tags related to glucose transporter GLUT1 of human erythrocyte. PiHXT5 is regulated in response to mutualism as well as glucose concentration. We have functionally characterized PiHXT5 by complementation of hxt-null mutant of Saccharomyces cerevisiae EBY.VW4000. It is involved in transport of multiple sugars ranging from D-glucose, D-fructose, D-xylose, Dmannose, D-galactose with decreasing affinity. The uncoupling experiments indicate that it functions as HC/glucose co-transporter. Further, pH dependence analysis suggests that it functions maximum between pH 5 and 6. The expression of PiHXT5 is dependent on glucose concentration and was found to be expressed at low glucose levels (1 mM) which indicate its role as a high affinity glucose transporter. Our study on this sugar transporter will help in better understanding of carbon metabolism and flow in this agro-friendly fungus. [ABSTRACT FROM AUTHOR]

Fission yeast, Schizosaccharomyces pombe, possesses eight hexose transporters, Ght1~8. In order to clarify the role of each hexose transporter on glucose uptake, a glucose uptake assay system was established and the actual glucose uptake activity of each hexose transporter-deletion mutant was measured. Under normal growth condition containing 2% glucose, ∆ght5 and ∆ght2 mutants showed large and small decrease in glucose uptake activity, respectively. On the other hand, the other deletion mutants did not show any decrease in glucose uptake activity indicating that, in the presence of Ght5 and Ght2, the other hexose transporters do not play a significant role in glucose uptake. To understand the relevance between glucose uptake and lifespan regulation, we measured the chronological lifespan of each hexose transporter deletion mutant, and found that only ∆ght5 mutant showed a significant lifespan extension. Based on these results we showed that Ght5 is mainly involved in the glucose uptake in Schizosaccharomyces pombe, and suggested that the ∆ght5 mutant has prolonged lifespan due to physiological changes similar to calorie restriction.

Background: A Saccharomyces cerevisiae strain carrying deletions in all three pyruvate decarboxylase (PDC) genes (also called Pdc negative yeast) represents a non-ethanol producing platform strain for the production of pyruvate derived biochemicals. However, it cannot grow on glucose as the sole carbon source, and requires supplementation of C2 compounds to the medium in order to meet the requirement for cytosolic acetyl-CoA for biosynthesis of fatty acids and ergosterol. Results: In this study, a Pdc negative strain was adaptively evolved for improved growth in glucose medium via serial transfer, resulting in three independently evolved strains, which were able to grow in minimal medium containing glucose as the sole carbon source at the maximum specific rates of 0.138, 0.148, 0.141 h-1, respectively. Several genetic changes were identified in the evolved Pdc negative strains by genomic DNA sequencing. Among these genetic changes, 4 genes were found to carry point mutations in at least two of the evolved strains: MTH1 encoding a negative regulator of the glucose-sensing signal transduction pathway, HXT2 encoding a hexose transporter, CIT1 encoding a mitochondrial citrate synthase, and RPD3 encoding a histone deacetylase. Reverse engineering of the non-evolved Pdc negative strain through introduction of the MTH181D allele restored its growth on glucose at a maximum specific rate of 0.053 h-1 in minimal medium with 2% glucose, and the CIT1 deletion in the reverse engineered strain further increased the maximum specific growth rate to 0.069 h-1. Conclusions: In this study, possible evolving mechanisms of Pdc negative strains on glucose were investigated by genome sequencing and reverse engineering. The non-synonymous mutations in MTH1 alleviated the glucose repression by repressing expression of several hexose transporter genes. The non-synonymous mutations in HXT2 and CIT1 may function in the presence of mutated MTH1 alleles and could be related to an altered central carbon metabolism in order to ensure production of cytosolic acetyl-CoA in the Pdc negative strain. [ABSTRACT FROM AUTHOR]

Many hexose transporters (HTs) have been reported to play roles in sucrose-transporting plants. However, little information about roles of HTs in RFOs (raffinose family oligosaccharides)-transporting plants has been reported. Here, three hexose transporters ( CsHT2 , CsHT3 , and CsHT4 ) were cloned from Cucumis sativus L. Heterologous expression in yeast demonstrated that CsHT3 transported glucose, galactose and mannose, with a K m of 131.9 μM for glucose, and CsHT4 only transported galactose, while CsHT2 was non-functional. Both CsHT3 and CsHT4 were targeted to the plasma membrane of cucumber protoplasts. Spatio-temporal expression indicated that transcript level of CsHT3 was much higher than that of CsHT2 and CsHT4 in most tissues, especially in peduncles and fruit tissues containing vascular bundles. GUS staining of CsHT3 -promoter-β-glucuronidase (GUS) transgenic Arabidopsis plants revealed CsHT3 expression in tissues with high metabolic turnover, suggesting that CsHT3 is involved in sugar competition among different sink organs during plant development. The transcript levels of CsHT3 and cell wall invertase genes increased in peduncles and fruit tissues along with cucumber fruit enlargement, and CsHT3 localized to phloem tissues by immunohistochemical localization; These results suggest that CsHT3 probably plays an important role in apoplastic phloem unloading of cucumber fruit. [ABSTRACT FROM AUTHOR]

Here we review cell cycle control in the fission yeast, Schizosaccharomyces pombe, in response to an abrupt reduction of glucose concentration in culture media. S. pombe arrests cell cycle progression when transferred from media containing 2.0% glucose to media containing 0.1%. After a delay, S. pombe resumes cell division at a surprisingly fast rate, comparable to that observed in 2% glucose. We found that a number of genes, including zinc-finger transcription factor Scr1, CaMKK-like protein kinase Ssp1, and glucose transporter Ght5, enable rapid cell division in low glucose. In this article, we examine whether cell cycle checkpoint-like control operates during the delay and after resumption of cell division in limited-glucose. Using microarray analysis and genetic screening, we identified several candidate genes that may be involved in controlling this low-glucose adaptation. [ABSTRACT FROM AUTHOR]

Highlights: [•] Overexpression of gene HXT13 confers resistance to miltefosine (MI) in S. cerevisiae. [•] Overexpression of similar genes HXT15 and HXT17 does not confer MI resistance. [•] MI treated HXT13 overexpressed cells had greater viability compared to control. [•] Hxt13p functions as an ATP-independent MI specific efflux pump. [•] HXT13 overexpressed strain was not multidrug resistant. [Copyright &y& Elsevier]

Schizosaccharomyces pombe has eight hexose transporter genes, ght1 to ght8. Here we report that ght2, which is highly expressed in the presence of glucose, is essential for UDP-galactose synthesis from extracellular galactose when cells grow on glucose. The galactosylation defect of a uge1Δ mutant defective in synthesis of UDP-galactose from glucose was suppressed in galactose-containing medium, but disruption of ght2 in the uge1Δ mutant reversed suppression of the galactosylation defect. Expression of Saccharomyces cerevisiae GAL2 in uge1Δ ght2Δ cells suppressed the defective galactosylation phenotype in galactose-containing medium. These results indicate that galactose is transported from the medium to the cytosol in a Ght2-dependent manner, and is then converted into UDP-galactose. [ABSTRACT FROM AUTHOR]

Mezcal from Tamaulipas (México) is produced by spontaneous alcoholic fermentation using Agave spp. musts, which are rich in fructose. In this study eight Saccharomyces cerevisiae isolates obtained at the final stage of fermentation from a traditional mezcal winery were analysed in three semi-synthetic media. Medium M1 had a sugar content of 100 g l and a glucose/fructose (G/F) of 9:1. Medium M2 had a sugar content of 100 g l and a G/F of 1:9. Medium M3 had a sugar content of 200 g l and a G/F of 1:1. In the three types of media tested, the highest ethanol yield was obtained from the glucophilic strain LCBG-3Y5, while strain LCBG-3Y8 was highly resistant to ethanol and the most fructophilic of the mezcal strains. Strain LCBG-3Y5 produced more glycerol (4.4 g l) and acetic acid (1 g l) in M2 than in M1 (1.7 and 0.5 g l, respectively), and the ethanol yields were higher for all strains in M1 except for LCBG-3Y5, -3Y8 and the Fermichamp strain. In medium M3, only the Fermichamp strain was able to fully consume the 100 g of fructose l but left a residual 32 g of glucose l. Regarding the hexose transporters, a high number of amino acid polymorphisms were found in the Hxt1p sequences. Strain LCBG-3Y8 exhibited eight unique amino acid changes, followed by the Fermichamp strain with three changes. In Hxt3p, we observed nine amino acid polymorphisms unique for the Fermichamp strain and five unique changes for the mezcal strains. [ABSTRACT FROM AUTHOR]

Despite intense elimination efforts, human malaria, caused by the infection of five Plasmodium species, remains the deadliest parasitic disease in the world. Even worse, with the emergence and spreading of the first-line drug-resistant Plasmodium parasites, therapeutic interventions based on novel plasmodial drug targets are more necessary than ever. Given that the blood-stage parasites primarily rely on glycolysis for their energy supply, blocking glucose uptake, the rate-limiting step of ATP generation, was considered a promising approach to kill these parasites. To achieve this goal, characterization of the plasmodial hexose transporter and development of selective inhibitors have been pursued for decades. Here, we review the identification and characterization of the Plasmodium falciparum hexose transporter (PfHT1) and summarize current advances in its inhibitor development., (© 2022 The Author. Proteins: Structure, Function, and Bioinformatics published by Wiley Periodicals LLC.)

Considerable amounts of information is available on the complex carbohydrates that are mobilized and utilized by the seed to support early seedling development. These events occur after radicle has protruded from the seed. However, scarce information is available on the role of the endogenous soluble carbohydrates from the embryo in the first hours of germination. The present work analysed how the soluble carbohydrate reserves in isolated maize embryos are mobilized during 6–24 h of water imbibition, an interval that exclusively embraces the first two phases of the germination process. It was found that sucrose constitutes a very significant reserve in the scutellum and that it is efficiently consumed during the time in which the adjacent embryo axis is engaged in an active metabolism. Sucrose transporter was immunolocalized in the scutellum and in vascular elements. In parallel, a cell-wall invertase activity, which hydrolyses sucrose, developed in the embryo axis, which favoured higher glucose uptake. Sucrose and hexose transporters were active in the embryo tissues, together with the plasma membrane H+-ATPase, which was localized in all embryo regions involved in both nutrient transport and active cell elongation to support radicle extension. It is proposed that, during the initial maize germination phases, a net flow of sucrose takes place from the scutellum towards the embryo axis and regions that undergo elongation. During radicle extension, sucrose and hexose transporters, as well as H+-ATPase, become the fundamental proteins that orchestrate the transport of nutrients required for successful germination. [ABSTRACT FROM AUTHOR]