Agrobacterium radiobacter and related organisms take up fructose via a binding-protein-dependent active-transport system

Williams, Steven G.; Greenwood, Jacqueline A.; Jones, Colin W.

doi:10.1099/13500872-141-10-2601

Cited by 7 publications

(9 citation statements)

References 20 publications

(22 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Basically, this transport was characterized as an active nonphosphorylating process (15), and more recently a periplasmic protein which is immunologically similar to an FBP from A. radiobacter had been detected (60). In this study, we have characterized the S. meliloti frc locus, which encodes a high-affinity transport system for fructose, mannose, and ribose.…”

Section: Discussionmentioning

confidence: 99%

“…Total cell proteins or periplasmic proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (PAGE) and transferred to nitrocellulose membrane (Hybond protein; 0.2-m pore size) by electroblotting. Immunoblotting was performed using a 1/5000 dilution of a polyclonal serum raised against the FBP of A. radiobacter (60). The immunoblots were developed with a Renaissance kit (Dupont/NEN) as instructed by the manufacturer.…”

Section: Methodsmentioning

confidence: 99%

“…In fact, fructose catabolism occurs by phosphorylation into fructose-6-phosphate and conversion into glucose-6-phosphate, prior to its metabolization via the Entner-Doudoroff pathway known to be present in these microorganisms (8,15,17). A periplasmic fructose-binding protein (FBP) has been detected recently in Agrobacterium radiobacter, a member of the family Rhizobiaceae, and Western blotting with antiserum to FBP has shown the presence of an immunologically similar protein in S. meliloti and R. leguminosarum (60). However, the structure of the fructose transport system has not been investigated.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Fructose Uptake in Sinorhizobium meliloti Is Mediated by a High-Affinity ATP-Binding Cassette Transport System

et al. 2001

View full text Add to dashboard Cite

By transposon mutagenesis, we have isolated a mutant of Sinorhizobium meliloti which is totally unable to grow on fructose as sole carbon source as a consequence of its inability to transport this sugar. The cloning and sequencing analysis of the chromosomal DNA region flanking the TnphoA insertion revealed the presence of six open reading frames (ORFs) organized in two loci, frcRS and frcBCAK, transcribed divergently. The frcBCA genes encode the characteristic components of an ATP-binding cassette transporter (FrcB, a periplasmic substrate binding protein, FrcC, an integral membrane permease, and FrcA, an ATP-binding cytoplasmic protein), which is the unique high-affinity (K m of 6 M) fructose uptake system in S. meliloti. The FrcK protein shows homology with some kinases, while FrcR is probably a transcriptional regulator of the repressor-ORFkinase family. The expression of S. meliloti frcBCAK in Escherichia coli, which transports fructose only via the phosphotransferase system, resulted in the detection of a periplasmic fructose binding activity, demonstrating that FrcB is the binding protein of the Frc transporter. The analysis of substrate specificities revealed that the Frc system is also a high-affinity transporter for ribose and mannose, which are both fructose competitors for the binding to the periplasmic FrcB protein. However, the Frc mutant was still able to grow on these sugars as sole carbon source, demonstrating the presence of at least one other uptake system for mannose and ribose in S. meliloti. The expression of the frcBC genes as determined by measurements of alkaline phosphatase activity was shown to be induced by mannitol and fructose, but not by mannose, ribose, glucose, or succinate, suggesting that the Frc system is primarily targeted towards fructose. Neither Nod nor Fix phenotypes were impared in the TnphoA mutant, demonstrating that fructose uptake is not essential for nodulation and nitrogen fixation, although FrcB protein is expressed in bacteroids isolated from alfalfa nodulated by S. meliloti wild-type strains.Carbohydrates represent one of the major structural building blocks of all organisms. In bacterial cells, three energydependent sugar uptake mechanisms have been characterized. One that is widely used operates by proton symport (19); this system belongs to the major facilitator superfamily MFS (43) and is used in Escherichia coli for the uptake of galactose, xylose, and lactose. A second system, the phosphoenolpyruvate:sugar phosphotransferase system (PTS), is found in many bacteria (44) and is the main transporter for glucose, fructose, mannose, and sucrose in many gram-negative bacteria (41). A third transport mechanism found in all three kingdoms is the periplasmic binding protein-dependent ATP-binding cassette (ABC)-type carrier (4). In bacteria, the ABC superfamily transports a wide range of substrates, including a variety of monosaccharides like arabinose or ribose as well as disaccharides such as maltose and tri-or higher oligosaccharides (43).Sinorhizobium meliloti i...

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Fructose Uptake in Sinorhizobium meliloti Is Mediated by a High-Affinity ATP-Binding Cassette Transport System

et al. 2001

View full text Add to dashboard Cite

show abstract

“…The first two systems are distantly related to other ABC carbohydrate transport systems and thus were predicted as putative porters for oligofructose. The third system is another ABC transporter ( fma , f ructose-ma nnose-like) with similarity to a fructose ABC permease of Agrobacterium radiobacter [Williams et al, 1995]. A glucose kinase gene (glkA) , a gene for an ATPase (fmaK) and a ROK family regulator are found upstream of fmaEFG .…”

Section: Abc Family: Lactose Raffinose Maltose and Fructooligosaccmentioning

confidence: 99%

Sugar Transport Systems of<i> Bifidobacterium longum </i>NCC2705

Parche

Amon²,

Janković³

et al. 2006

Microb Physiol

101

View full text Add to dashboard Cite

Here we present the complement of the carbohydrate uptake systems of the strictly anaerobic probiotic Bifidobacterium longum NCC2705. The genome analysis of this bacterium predicts that it has 19 permeases for the uptake of diverse carbohydrates. The majority belongs to the ATP-binding cassette transporter family with 13 systems identified. Among them are permeases for lactose, maltose, raffinose, and fructooligosaccharides, a commonly used prebiotic additive. We found genes that encode a complete phosphotransferase system (PTS) and genes for three permeases of the major facilitator superfamily. These systems could serve for the import of glucose, galactose, lactose, and sucrose. Growth analysis of NCC2705 cells combined with biochemical characterization and microarray data showed that the predicted substrates are consumed and that the corresponding transport and catabolic genes are expressed. Biochemical analysis of the PTS, in which proteins are central in regulation of carbon metabolism in many bacteria, revealed that B. longum has a glucose-specific PTS, while two other species (Bifidobacterium lactis and Bifidobacterium bifidum) have a fructose-6-phosphate-forming fructose-PTS instead. It became obvious that most carbohydrate systems are closely related to those from other actinomycetes, with a few exceptions. We hope that this report on B. longum carbohydrate transporter systems will serve as a guide for further in-depth analyses on the nutritional lifestyle of this beneficial bacterium.

show abstract

“…Similar systems for the uptake of octopine (40), nopaline (47), and agrocinopine B (3,18) and the export of ␤-1,2-glucan (7) have been reported for A. tumefaciens. Lactose (17,43), fructose (42), galactose, and glucose (11) transporters have been described for A. radiobacter, an avirulent species of Agrobacterium. A. radiobacter was found to contain two periplasmic high-affinity glucosebinding proteins, GBP1 and GBP2 (11).…”

mentioning

confidence: 99%

The Agrobacterium tumefaciens virulence gene chvE is part of a putative ABC-type sugar transport operon

1997

View full text Add to dashboard Cite

The Agrobacterium tumefaciens virulence determinant ChvE is a periplasmic binding protein which participates in chemotaxis and virulence gene induction in response to monosaccharides which occur in the plant wound environment. The region downstream of the A. tumefaciens chvE gene was cloned and sequenced for nucleotide and expression analysis. Three open reading frames transcribed in the same direction as chvE were revealed. The first two, together with chvE, encode putative proteins of a periplasmic binding protein-dependent sugar uptake system, or ABC-type (ATP binding cassette) transporter. The third open reading frame encodes a protein of unknown function. The deduced transporter gene products are related on the amino acid level to bacterial sugar transporters and probably function in glucose and galactose uptake. We have named these genes gguA, -B, and -C, for glucose galactose uptake. Mutations in gguA, gguB, or gguC do not affect virulence of A. tumefaciens on Kalanchöe diagremontiana; growth on 1 mM galactose, glucose, xylose, ribose, arabinose, fucose, or sucrose; or chemotaxis toward glucose, galactose, xylose, or arabinose.Agrobacterium tumefaciens is a gram-negative bacterial pathogen that forms crown gall tumors on dicotyledonous plants by transferring a piece of its DNA into the plant cell, where it becomes stably integrated into the host genome (for a review, see reference 44). The transferred DNA, termed the T-DNA, is processed and transported from the bacterial cell by components encoded by the vir regulon. The vir genes are induced in response to chemical signals of the plant wound site, which include low pH, phenolic compounds, and monosaccharide components of the plant cell wall (6,33,38,39). Monosaccharides are bound by the periplasmic sugar-binding protein ChvE, which then interacts with the periplasmic region of the membrane-bound VirA molecule of the VirA-VirG sensor-regulator pair to activate transcription of the vir regulon (24,34,45). ChvE also mediates chemotaxis toward various sugars, presumably by interacting in the sugar-bound form with an unidentified membrane-bound receptor (6). Thus, this sugar-binding protein plays at least two roles in the physiology and virulence of A. tumefaciens.ChvE may have an additional role as well. In gram-negative bacteria, periplasmic binding proteins like ChvE often constitute part of a high-affinity uptake system (22, 23). These transporters couple hydrolysis of ATP to transport and are capable of taking up solutes at low concentrations (micromolar range and less) and concentrating them across a large gradient. Structurally, they are composed of a minimum of three components: a periplasmic binding protein, a membrane-bound transport protein(s) which interacts with the periplasmic protein bound to its ligand, and an ATP-binding protein(s) which energizes transport at the cytoplasmic face of the membrane. These periplasmic binding protein-dependent transporters take up a wide assortment of solutes, and a large number of separate transport systems for...

show abstract

Agrobacterium radiobacter and related organisms take up fructose via a binding-protein-dependent active-transport system

Cited by 7 publications

References 20 publications

Fructose Uptake in Sinorhizobium meliloti Is Mediated by a High-Affinity ATP-Binding Cassette Transport System

Fructose Uptake in Sinorhizobium meliloti Is Mediated by a High-Affinity ATP-Binding Cassette Transport System

Sugar Transport Systems of<i> Bifidobacterium longum </i>NCC2705

The Agrobacterium tumefaciens virulence gene chvE is part of a putative ABC-type sugar transport operon

Contact Info

Product

Resources

About