scholarly journals Genome-Wide Analysis of Sugar Transporters Identifies the gtsA Gene for Glucose Transportation in Pseudomonas stutzeri A1501

2020 ◽  
Vol 8 (4) ◽  
pp. 592
Author(s):  
Yaqun Liu ◽  
Liguo Shang ◽  
Yuhua Zhan ◽  
Min Lin ◽  
Zhu Liu ◽  
...  
Keyword(s):  
Gene Diversity ◽  
Carbon Sources ◽  
Sugar Transport ◽  
Genomic Analysis ◽  
Pseudomonas Stutzeri ◽  
Major Facilitator Superfamily ◽  
Transport Systems ◽  
Phosphotransferase System ◽  
Metabolic System ◽  
Sugar Transporters

Pseudomonas stutzeri A1501 possesses an extraordinary number of transporters which confer this rhizosphere bacterium with the sophisticated ability to metabolize various carbon sources. However, sugars are not a preferred carbon source for P. stutzeri A1501. The P. stutzeri A1501 genome has been sequenced, allowing for the homology-based in silico identification of genes potentially encoding sugar-transport systems by using established microbial sugar transporters as a template sequence. Genomic analysis revealed that there were 10 sugar transporters in P. stutzeri A1501, most of which belong to the ATP-binding cassette (ABC) family (5/10); the others belong to the phosphotransferase system (PTS), major intrinsic protein (MIP) family, major facilitator superfamily (MFS) and the sodium solute superfamily (SSS). These systems might serve for the import of glucose, galactose, fructose and other types of sugar. Growth analysis showed that the only effective medium was glucose and its corresponding metabolic system was relatively complete. Notably, the loci of glucose metabolism regulatory systems HexR, GltR/GtrS, and GntR were adjacent to the transporters ABCMalEFGK, ABCGtsABCD, and ABCMtlEFGK, respectively. Only the ABCGtsABCD expression was significantly upregulated under both glucose-sufficient and -limited conditions. The predicted structure and mutant phenotype data of the key protein GtsA provided biochemical evidence that P. stutzeri A1501 predominantly utilized the ABCGtsABCD transporter for glucose uptake. We speculate that gene absence and gene diversity in P. stutzeri A1501 was caused by sugar-deficient environmental factors and hope that this report can provide guidance for further analysis of similar bacterial lifestyles.

scholarly journals A Genomic View of Sugar Transport in Mycobacterium smegmatis and Mycobacterium tuberculosis

2007 ◽  
Vol 189 (16) ◽  
pp. 5903-5915 ◽  
Author(s):  
Fritz Titgemeyer ◽  
Johannes Amon ◽  
Stephan Parche ◽  
Maysa Mahfoud ◽  
Johannes Bail ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Mycobacterium Smegmatis ◽  
Glucose Transporter ◽  
Sugar Transport ◽  
Major Facilitator Superfamily ◽  
Transport Systems ◽  
Phosphotransferase System ◽  
Natural Habitats ◽  
Transporter Family ◽  
Slow Growing

ABSTRACT We present a comprehensive analysis of carbohydrate uptake systems of the soil bacterium Mycobacterium smegmatis and the human pathogen Mycobacterium tuberculosis. Our results show that M. smegmatis has 28 putative carbohydrate transporters. The majority of sugar transport systems (19/28) in M. smegmatis belong to the ATP-binding cassette (ABC) transporter family. In contrast to previous reports, we identified genes encoding all components of the phosphotransferase system (PTS), including permeases for fructose, glucose, and dihydroxyacetone, in M. smegmatis. It is anticipated that the PTS of M. smegmatis plays an important role in the global control of carbon metabolism similar to those of other bacteria. M. smegmatis further possesses one putative glycerol facilitator of the major intrinsic protein family, four sugar permeases of the major facilitator superfamily, one of which was assigned as a glucose transporter, and one galactose permease of the sodium solute superfamily. Our predictions were validated by gene expression, growth, and sugar transport analyses. Strikingly, we detected only five sugar permeases in the slow-growing species M. tuberculosis, two of which occur in M. smegmatis. Genes for a PTS are missing in M. tuberculosis. Our analysis thus brings the diversity of carbohydrate uptake systems of fast- and a slow-growing mycobacteria to light, which reflects the lifestyles of M. smegmatis and M. tuberculosis in their natural habitats, the soil and the human body, respectively.

scholarly journals Aspergillus niger mstA encodes a high-affinity sugar/H+ symporter which is regulated in response to extracellular pH

2004 ◽  
Vol 379 (2) ◽  
pp. 375-383 ◽  
Author(s):  
Patricia A. vanKUYK ◽  
Jasper A. DIDERICH ◽  
Andrew P. MacCABE ◽  
Oscar HERERRO ◽  
George J. G. RUIJTER ◽  
...  
Keyword(s):  
Aspergillus Niger ◽  
Ph Regulation ◽  
Functional Characterization ◽  
Carbon Sources ◽  
Sugar Transport ◽  
Major Facilitator Superfamily ◽  
Northern Analysis ◽  
High Affinity ◽  
Hexose Uptake ◽  
Major Facilitator

A sugar-transporter-encoding gene, mstA, which is a member of the major facilitator superfamily, has been cloned from a genomic DNA library of the filamentous fungus Aspergillus niger. To enable the functional characterization of MSTA, a full-length cDNA was expressed in a Saccharomyces cerevisiae strain deficient in hexose uptake. Uptake experiments using 14C-labelled monosaccharides demonstrated that although able to transport d-fructose (Km, 4.5±1.0 mM), d-xylose (Km, 0.3±0.1 mM) and d-mannose (Km, 60±20 µM), MSTA has a preference for d-glucose (Km, 25±10 µM). pH changes associated with sugar transport indicate that MSTA catalyses monosaccharide/H+ symport. Expression of mstA in response to carbon starvation and upon transfer to poor carbon sources is consistent with a role for MSTA as a high-affinity transporter for d-glucose, d-mannose and d-xylose. Northern analysis has shown that mstA is subject to CreA-mediated carbon catabolite repression and pH regulation mediated by PacC. A. niger strains in which the mstA gene had been disrupted are phenotypically identical with isogenic reference strains when grown on 0.1–60 mM d-glucose, d-mannose, d-fructose or d-xylose. This indicates that A. niger possesses other transporters capable of compensating for the absence of MSTA.

scholarly journals In Silico and Transcriptional Analysis of Carbohydrate Uptake Systems of Streptomyces coelicolor A3(2)

2004 ◽  
Vol 186 (5) ◽  
pp. 1362-1373 ◽  
Author(s):  
Ralph Bertram ◽  
Maximilian Schlicht ◽  
Kerstin Mahr ◽  
Harald Nothaft ◽  
Milton H. Saier ◽  
...  
Keyword(s):  
Antibiotic Production ◽  
Streptomyces Coelicolor ◽  
Essential Feature ◽  
Carbon Sources ◽  
Gene Clusters ◽  
Major Facilitator Superfamily ◽  
Transcriptional Analysis ◽  
Phosphotransferase System ◽  
Function Similarity ◽  
Major Facilitator

ABSTRACT Streptomyces coelicolor is the prototype for the investigation of antibiotic-producing and differentiating actinomycetes. As soil bacteria, streptomycetes can metabolize a wide variety of carbon sources and are hence vested with various specific permeases. Their activity and regulation substantially determine the nutritional state of the cell and, therefore, influence morphogenesis and antibiotic production. We have surveyed the genome of S. coelicolor A3(2) to provide a thorough description of the carbohydrate uptake systems. Among 81 ATP-binding cassette (ABC) permeases that are present in the genome, we found 45 to encode a putative solute binding protein, an essential feature for carbohydrate permease function. Similarity analysis allowed the prediction of putative ABC systems for transport of cellobiose and cellotriose, α-glucosides, lactose, maltose, maltodextrins, ribose, sugar alcohols, xylose, and β-xylosides. A novel putative bifunctional protein composed of a substrate binding and a membrane-spanning moiety is likely to account for ribose or ribonucleoside uptake. Glucose may be incorporated by a proton-driven symporter of the major facilitator superfamily while a putative sodium-dependent permease of the solute-sodium symporter family may mediate uptake of galactose and a facilitator protein of the major intrinsic protein family may internalize glycerol. Of the predicted gene clusters, reverse transcriptase PCRs showed active gene expression in 8 of 11 systems. Together with the previously surveyed permeases of the phosphotransferase system that accounts for the uptake of fructose and N-acetylglucosamine, the genome of S. coelicolor encodes at least 53 potential carbohydrate uptake systems.

The regulatory link between carbon and nitrogen metabolism in Bacillus subtilis: regulation of the gltAB operon by the catabolite control protein CcpA

Microbiology ◽  
2003 ◽  
Vol 149 (10) ◽  
pp. 3001-3009 ◽  
Author(s):  
Ingrid Wacker ◽  
Holger Ludwig ◽  
Irene Reif ◽  
Hans-Matti Blencke ◽  
Christian Detsch ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Carbon Sources ◽  
Sugar Transport ◽  
Glutamate Synthase ◽  
Phosphotransferase System ◽  
Carbon And Nitrogen ◽  
Artificial Induction ◽  
Key Factor ◽  
Regulatory Link ◽  
Control Protein

Bacillus subtilis assimilates ammonium by the concerted action of glutamine synthetase and glutamate synthase. The expression of the gltAB operon encoding the latter enzyme is impaired in B. subtilis ccpA mutant strains. CcpA is a pleiotropic transcriptional regulator that is the key factor in the regulation of carbon metabolism. However, in addition to their defect in catabolite repression ccpA mutants are unable to grow on minimal media with glucose and ammonium as the single sources of carbon and nitrogen, respectively. In this work, the expression of the gltAB operon was analysed and its role in growth on minimal sugar/ammonium media was studied. Expression of gltAB requires induction by glucose or other glycolytically catabolized carbon sources. In ccpA mutants, gltAB cannot be induced by glucose due to the low activity of the phosphotransferase sugar transport system in these mutants. A mutation that allowed phosphotransferase system activity in a ccpA background simultaneously restored glucose induction of gltAB and growth on glucose/ammonium medium. Moreover, artificial induction of the gltAB operon in the ccpA mutant allowed the mutant strain to grow on minimal medium with glucose and ammonium. It may be concluded that expression of the gltAB operon depends on the accumulation of glycolytic intermediates which cannot occur in the ccpA mutant. The lack of gltAB induction is the bottleneck that prevents growth of the ccpA mutant on glucose/ammonium media. The control of expression of the gltAB operon by CcpA provides a major regulatory link between carbon and amino acid metabolism.

scholarly journals Effects of Homologous Phosphoenolpyruvate-Carbohydrate Phosphotransferase System Proteins on Carbohydrate Uptake and Poly(3-Hydroxybutyrate) Accumulation in Ralstonia eutropha H16

2011 ◽  
Vol 77 (11) ◽  
pp. 3582-3590 ◽  
Author(s):  
Chlud Kaddor ◽  
Alexander Steinbüchel
Keyword(s):  
Ralstonia Eutropha ◽  
Insertion Site ◽  
Carbon Sources ◽  
Sugar Transport ◽  
Dry Weight ◽  
Wild Type ◽  
Phosphotransferase System ◽  
Homologous Proteins ◽  
Content Type ◽  
Negative Effect

ABSTRACTSeven gene loci encoding putative proteins of the phosphoenolpyruvate-carbohydrate phosphotransferase system (PEP-PTS) were identified in the genome ofRalstonia eutrophaH16 byin silicoanalysis. Except theN-acetylglucosamine-specific PEP-PTS, an additional complete PEP-PTS is lacking in strain H16. Based on these findings, we generated single and multiple deletion mutants defective mainly in the PEP-PTS genes to investigate their influence on carbon source utilization, growth behavior, and poly(3-hydroxybutyrate) (PHB) accumulation. As supposed, the H16 ΔfrcACBand H16 ΔnagFECmutants exhibited no growth when cultivated on fructose andN-acetylglucosamine, respectively. Furthermore, a transposon mutant with aptsM-ptsHinsertion site did not grow on both carbon sources. The observed phenotype was not complemented, suggesting that it results from an interaction of genes or a polar effect caused by the Tn5::mobinsertion.ptsM,ptsH, andptsIsingle, double, and triple mutants stored much less PHB than the wild type (about 10 to 39% [wt/wt] of cell dry weight) and caused reduced PHB production in mutants lacking the H16_A2203, H16_A0384,frcACB, ornagFECgenes. In contrast, mutant H16 ΔH16_A0384 accumulated 11.5% (wt/wt) more PHB than the wild type when grown on gluconate and suppressed partially the negative effect of theptsMHIdeletion on PHB synthesis. Based on our experimental data, we discussed whether the PEP-PTS homologous proteins inR. eutrophaH16 are exclusively involved in the complex sugar transport system or whether they are also involved in cellular regulatory functions of carbon and PHB metabolism.

The osmotolerant fructophilic yeast Zygosaccharomyces rouxii employs two plasma-membrane fructose uptake systems belonging to a new family of yeast sugar transporters

Microbiology ◽  
2011 ◽  
Vol 157 (2) ◽  
pp. 601-608 ◽  
Author(s):  
Maria José Leandro ◽  
Hana Sychrová ◽  
Catarina Prista ◽  
Maria C. Loureiro-Dias
Keyword(s):  
Plasma Membrane ◽  
Diffusion Mechanism ◽  
Sugar Transport ◽  
High Capacity ◽  
Transport Systems ◽  
Facilitated Diffusion ◽  
Sugar Transporters ◽  
Zygosaccharomyces Rouxii ◽  
New Family ◽  
Spoilage Yeasts

Owing to its high resistance to weak-acid preservatives and extreme osmotolerance, Zygosaccharomyces rouxii is one of the main spoilage yeasts of sweet foods and beverages. In contrast with Saccharomyces cerevisiae, Z. rouxii is a fructophilic yeast; it consumes fructose faster than glucose. So far, to our knowledge, no specific Z. rouxii proteins responsible for this fructophilic behaviour have been characterized. We have identified two genes encoding putative fructose transporters in the Z. rouxii CBS 732 genome. Heterologous expression of these two Z. rouxii ORFs in a S. cerevisiae strain lacking its own hexose transporters (hxt-null) and subsequent kinetic analysis of sugar transport showed that both proteins are functionally expressed at the plasma membrane: ZrFfz1 is a high-capacity fructose-specific facilitator (K m∼400 mM and V max∼13 mmol h−1 g−1) and ZrFfz2 is a facilitator transporting glucose and fructose with similar capacity and affinity (K m∼200 mM and V max∼4 mmol h−1 g−1). These two proteins together with the Zygosaccharomyces bailii Ffz1 fructose-specific transporter belong to a new family of sugar transport systems mediating the uptake of hexoses via the facilitated diffusion mechanism, and are more homologous to drug/H+ antiporters (regarding their primary protein structure) than to other yeast sugar transporters of the Sugar Porter family.

scholarly journals Protein:Protein Interactions in the Cytoplasmic Membrane Influencing Sugar Transport and Phosphorylation Activities of theE. coliPhosphotransferase System

2019 ◽  
Author(s):  
Mohammad Aboulwafa ◽  
Zhongge Zhang ◽  
Milton H. Saier
Keyword(s):  
Protein Interactions ◽  
Cytoplasmic Membrane ◽  
Molecular Genetic ◽  
Sugar Transport ◽  
Specific Protein ◽  
Transport Systems ◽  
Phosphotransferase System ◽  
Protein Levels

AbstractThe multicomponent phosphoenolpyruvate-dependent sugar-transporting phosphotransferase system (PTS) inEscherichia colitakes up sugar substrates and concomitantly phosphorylates them. We have recently provided evidence that many of the integral membrane PTS permeases interact with the fructose PTS (FruA/FruB) [1]. However, the biochemical and physiological significance of this finding was not known. We have carried out molecular genetic/biochemical/physiological studies that show that interactions of the fructose PTS often enhance, but sometimes inhibit the activities of other PTS transporters many fold, depending on the target PTS system under study. Thus, the glucose, mannose, mannitol and N- acetylglucosamine permeases exhibit enhancedin vivosugar transport and sometimesin vitroPEP-dependent sugar phosphorylation activities while the galactitol and trehalose systems show inhibited activities. This is observed when the fructose system is induced to high levels and prevented when thefruA/fruBgenes are deleted. Overexpression of thefruAand/orfruBgenes in the absence of fructose induction during growth also enhances the rates of uptake of other hexoses. The β-galactosidase activities ofman, mtl,andgat-lacZtranscriptional fusions and the sugar-specific transphosphorylation activities of these enzyme transporters were not affected either by frustose induction orfruABoverexpression, showing that the rates of synthesis and protein levels in the membrane of the target PTS permeases were not altered. We thus suggest that specific protein-protein interactions within the cytoplasmic membrane regulate transportin vivo(and sometimes the PEP-dependent phosphorylation activitiesin vitroof PTS permeases) in a physiologically meaningful way that may help to provide a hierarchy of preferred PTS sugars. These observations appear to be applicable in principle to other types of transport systems as well.

scholarly journals SgrT, a Small Protein That Packs a Sweet Punch

2017 ◽  
Vol 199 (11) ◽  
Author(s):  
Medha Raina ◽  
Gisela Storz
Keyword(s):  
Amino Acid ◽  
Small Rna ◽  
Carbon Sources ◽  
Sugar Transport ◽  
Phosphotransferase System ◽  
Small Protein ◽  
Glucose Phosphate ◽  
Acid Protein ◽  
The Stability ◽  
Amino Acid Protein

ABSTRACT The SgrS small RNA (sRNA) has been shown to protect against elevated levels of glucose phosphate by regulating the stability and translation of mRNAs encoding proteins involved in sugar transport and catabolism. The sRNA also was known to encode a protective 43-amino-acid protein, SgrT, but little was known about its mechanism of action. Lloyd et al. (J Bacteriol 199:e00869-16, 2017, https://doi.org/10.1128/JB.00869-16 ) use cell biological and genetic approaches to demonstrate that the small protein interacts with the PtsG importer to block glucose transport by this phosphotransferase system and promote utilization of nonpreferred carbon sources to maintain growth during glucose-phosphate stress.

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