Anaerobic growth of Escherichia coli on d-tartrate depends on the fumarate carrier DcuB and fumarase, rather than the l-tartrate carrier TtdT and l-tartrate dehydratase

Kim, Ok Bin; Lux, Sebastián; Unden, Gottfried

doi:10.1007/s00203-007-0279-9

Cited by 29 publications

(21 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For genetic experiments, the bacteria were routinely grown in Luria-Bertani (LB) broth and on LB agar (19). For anaerobic expression studies, M9 medium or enriched M9 (eM9) medium (20,21), with acid-hydrolyzed casamino acids (0.1%) and tryptophan (0.005%), was used. Glucose (20 mM), glycerol (50 mM), DMSO (50 mM), and sodium fumarate (50 mM) were added as energy substrates, as indicated.…”

Section: Methodsmentioning

confidence: 99%

The Fumarate/Succinate Antiporter DcuB of Escherichia coli Is a Bifunctional Protein with Sites for Regulation of DcuS-dependent Gene Expression

Kleefeld

Ackermann

Bauer

et al. 2009

Journal of Biological Chemistry

Self Cite

103

View full text Add to dashboard Cite

DcuB of Escherichia coli catalyzes C 4 -dicarboxylate/succinate antiport during growth by fumarate respiration. The expression of genes of fumarate respiration, including the genes for DcuB (dcuB) and fumarate reductase (frdABCD) is transcriptionally activated by C 4 -dicarboxylates via the DcuS-DcuR two-component system, comprising the sensor kinase DcuS, which contains a periplasmic sensing domain for C 4 -dicarboxylates. Deletion or inactivation of dcuB caused constitutive expression of DcuSregulated genes in the absence of C 4 -dicarboxylates. The effect was specific for DcuB and not observed after inactivation of the homologous DcuA or the more distantly related DcuC transporter. Random and site-directed mutation identified three point mutations (T394I, D398N, and K353A) in DcuB that caused a similar derepression as dcuB deletion, whereas the transport activity of the DcuB mutants was retained. Constitutive expression in the dcuB mutants depended on the presence of a functional DcuS-DcuR two-component system. Mutation of residues E79A, R83A, and R127A of DcuB, on the other hand, inactivated growth by fumarate respiration and transport of [ 14 C]succinate, whereas the expression of dcuBlacZ was not affected. Therefore, the antiporter DcuB is a bifunctional protein and has a regulatory function that is independent from transport, and sites for transport and regulation can be differentiated.The fumarate/succinate antiporter DcuB (dicarboxylate uptake) of Escherichia coli catalyzes the uptake of external C 4 -dicarboxylates like fumarate, L-malate, or aspartate (1-3). Fumarate is used as an electron acceptor in fumarate respiration by fumarate reductase, which carries the active site at the cytoplasmic side of the membrane (for reviews, see Refs. 4 -6). L-Malate and aspartate are converted to fumarate by fumarase and aspartase, respectively, and are then metabolized in the same way as fumarate. Fumarate respiration results in the generation of a proton potential and drives ATP synthesis and growth of the bacteria. The product of fumarate respiration, succinate, is not further catabolized in anaerobic growth and is excreted by DcuB. DcuB, therefore, is responsible for the substrate/product antiport of fumarate, L-malate, or aspartate against succinate. DcuB and fumarase B are encoded by the dcuB fumB gene cluster (1, 7).

show abstract

Section: Methodsmentioning

confidence: 99%

The Fumarate/Succinate Antiporter DcuB of Escherichia coli Is a Bifunctional Protein with Sites for Regulation of DcuS-dependent Gene Expression

Kleefeld

Ackermann

Bauer

et al. 2009

Journal of Biological Chemistry

Self Cite

103

View full text Add to dashboard Cite

show abstract

“…thermopropionicum fumarase MmcBC, but several other fumarases have been described to be able to catalyse the dehydratation of the stereoisomer D-tartrate, but not L-tartrate or meso-tartrate, to oxaloacetate (Flint, 1994;Kim et al, 2007;Nakamura & Ogata, 1967, 1968a.…”

Section: Introductionmentioning

confidence: 99%

Identification of two [4Fe–4S]-cluster-containing hydro-lyases from Pyrococcus furiosus

et al. 2009

View full text Add to dashboard Cite

The hyperthermophilic archaeon Pyrococcus furiosus is a strict anaerobe. It is therefore not expected to use the oxidative tricarboxylic acid (TCA) cycle for energy transduction. Nonetheless, its genome encodes more putative TCA cycle enzymes than the closely related Pyrococcus horikoshii and Pyrococcus abyssi, including an aconitase (PF0201). Furthermore, a two-subunit fumarase (PF1755 and PF1754) is encoded on the Pyr. furiosus genome. In the present study, these three genes were heterologously overexpressed in Escherichia coli to enable characterization of the enzymes. PF1755 and PF1754 were shown to form a [4Fe-4S]-clustercontaining heterodimeric enzyme, able to catalyse the reversible hydratation of fumarate. The aconitase PF0201 also contained an Fe-S cluster, and catalysed the conversion from citrate to isocitrate. The fumarase belongs to the class of two-subunit, [4Fe-4S]-cluster-containing fumarate hydratases exemplified by MmcBC from Pelotomaculum thermopropionicum; the aconitase belongs to the aconitase A family. Aconitase probably plays a role in amino acid synthesis when the organism grows on carbohydrates. However, the function of the seemingly metabolically isolated fumarase in Pyr. furiosus has yet to be established.

show abstract

“…Because of the high selectivity of this SLC26 transporter for fumarate, a designation of FumT may be appropriate. In E. coli, fumarate uptake is mediated by the unrelated DcuB, a C4-dicarboxylate/ succinate antiporter active during growth by fumarate respiration 20 .…”

Section: Reinhart a F Reithmeier And Trevor F Moraesmentioning

confidence: 99%

Solute carriers keep on rockin'

Reithmeier

Moraes

2015

Nat Struct Mol Biol

View full text Add to dashboard Cite

Anaerobic growth of Escherichia coli on d-tartrate depends on the fumarate carrier DcuB and fumarase, rather than the l-tartrate carrier TtdT and l-tartrate dehydratase

Cited by 29 publications

References 37 publications

The Fumarate/Succinate Antiporter DcuB of Escherichia coli Is a Bifunctional Protein with Sites for Regulation of DcuS-dependent Gene Expression

The Fumarate/Succinate Antiporter DcuB of Escherichia coli Is a Bifunctional Protein with Sites for Regulation of DcuS-dependent Gene Expression

Identification of two [4Fe–4S]-cluster-containing hydro-lyases from Pyrococcus furiosus

Solute carriers keep on rockin'

Contact Info

Product

Resources

About