Partial Replacement of Succinate Dehydrogenase Function by Phage- and Plasmid-specified Fumarate Reductase in Escherichia coli

Guest, John R.

doi:10.1099/00221287-122-2-171

Cited by 50 publications

(58 citation statements)

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“…Under these conditions, a functional FRD is required; i.e., E. coli DW35 does not grow in either of these media. FRD when expressed from multicopy plasmids is fully capable of functionally replacing SDH in E. coli (13). FRD when expressed has, in fact, the same turnover number for succinate oxidation as native E. coli SDH (2, 33).…”

Section: Resultsmentioning

confidence: 99%

“…E. coli FRD catalyzes succinate oxidation at rates similar to that of the native SDH of that organism (2) and at about 30 to 40% of the rate at which it reduces fumarate (8). The frd gene products will replace SDH in sdh mutants when expressed from multicopy plasmids (13). It has been proposed, on the basis of sequence comparison of the catalytic domain subunits of FRD and SDH, that the genes are of common ancestral origin (41).…”

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confidence: 99%

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Aerobic inactivation of fumarate reductase from Escherichia coli by mutation of the [3Fe-4S]-quinone binding domain

Cecchini¹,

Sices²,

Schröder³

et al. 1995

J Bacteriol

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Fumarate reductase from Escherichia coli functions both as an anaerobic fumarate reductase and as an aerobic succinate dehydrogenase. A site-directed mutation of E. coli fumarate reductase in which FrdB Pro-159 was replaced with a glutamine or histidine residue was constructed and overexpressed in a strain of E. coli lacking a functional copy of the fumarate reductase or succinate dehydrogenase complex. The consequences of these mutations on bacterial growth, assembly of the enzyme complex, and enzymatic activity were investigated. Both mutations were found to have no effect on anaerobic bacterial growth or on the ability of the enzyme to reduce fumarate compared with the wild-type enzyme. The FrdB Pro-159-to-histidine substitution was normal in its ability to oxidize succinate. In contrast, however, the FrdB Pro-159-to-Gln substitution was found to inhibit aerobic growth of E. coli under conditions requiring a functional succinate dehydrogenase, and furthermore, the aerobic activity of the enzyme was severely inhibited upon incubation in the presence of its substrate, succinate. This inactivation could be prevented by incubating the mutant enzyme complex in an anaerobic environment, separating the catalytic subunits of the fumarate reductase complex from their membrane anchors, or blocking the transfer of electrons from the enzyme to quinones. The results of these studies suggest that the succinate-induced inactivation occurs by the production of hydroxyl radicals generated by a Fenton-type reaction following introduction of this mutation into the [3Fe-4S] binding domain. Additional evidence shows that the substrate-induced inactivation requires quinones, which are the membrane-bound electron acceptors and donors for the succinate dehydrogenase and fumarate reductase activities. These data suggest that the [3Fe-4S] cluster is intimately associated with one of the quinone binding sites found in fumarate reductase and succinate dehydrogenase.During anaerobic respiration in Escherichia coli, the fumarate reductase (FRD) complex catalyzes the fumarate-dependent oxidation of menaquinol, thereby allowing production of a transmembrane proton gradient that can be used by the organism to support metabolism (19). The FRD complex is composed of four nonidentical subunits, FrdA, FrdB, FrdC, and FrdD, arranged into two domains: the FrdAB catalytic domain and the FrdCD membrane anchor domain (2, 22). FrdA (66 kDa) contains covalently bound flavin adenine dinucleotide and the active site of the enzyme (38), and the 27-kDa FrdB subunit contains the three distinct iron-sulfur centers present in the enzyme (23). The iron-sulfur clusters, center 1 ([2Fe-2S] ), are similar in structure and function in all membranebound succinate dehydrogenases (SDHs) and FRDs examined to date (2). The FrdC (15 kDa) and FrdD (13 kDa) subunits are essential for attachment of the catalytic subunits to the inner surface of the cytoplasmic membrane (22) and are also essential for electron transfer reactions involving menaquinol and ubiquinone (8).The p...

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Section: Resultsmentioning

confidence: 99%

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confidence: 99%

Aerobic inactivation of fumarate reductase from Escherichia coli by mutation of the [3Fe-4S]-quinone binding domain

Cecchini¹,

Sices²,

Schröder³

et al. 1995

J Bacteriol

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“…However, their experiments did show activity for fumarate reductase. Evidence was presented previously which showed that fumarate reductase can compensate for a loss of succinate dehydrogenase (35,36). This could putatively convert succinate to fumarate.…”

Section: Figmentioning

confidence: 99%

Functional γ-Aminobutyrate Shunt in Listeria monocytogenes: Role in Acid Tolerance and Succinate Biosynthesis

Feehily

O’Byrne

Karatzas

2013

Appl Environ Microbiol

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Listeria monocytogenes, the causative agent of human listeriosis, is known for its ability to withstand severe environmental stresses. The glutamate decarboxylase (GAD) system is one of the principal systems utilized by the bacterium to cope with acid stress, a reaction that produces ␥-aminobutyrate (GABA) from glutamate. Recently, we have shown that GABA can accumulate intracellularly under acidic conditions, even under conditions where no extracellular glutamate-GABA exchange is detectable. The GABA shunt, a pathway that metabolizes GABA to succinate, has been described for several other bacterial genera, and the present study sought to determine whether L. monocytogenes has this metabolic capacity, which, if present, could provide a possible route for succinate biosynthesis in L. monocytogenes. Using crude protein extracts from L. monocytogenes EGD-e, we show that this strain exhibits activity for the two main enzyme reactions in the GABA shunt, GABA aminotransferase (GABA-AT) and succinic semialdehyde dehydrogenase (SSDH). Two genes were identified as candidates for encoding these enzyme activities, argD (GABA-AT) and lmo0913 (SSDH). Crude protein extracts prepared from a mutant lacking a functional argD gene significantly reduced GABA-AT activity, while an lmo0913 mutant lost all detectable SSDH activity. The deletion of lmo0913 increased the acid tolerance of EGD-e and showed an increased accumulation of intracellular GABA, suggesting that this pathway plays a significant role in the survival of this pathogen under acidic conditions. This is the first report of such a pathway in the genus Listeria, which highlights an important link between metabolism and acid tolerance and also presents a possible compensatory pathway to partially overcome the incomplete tricarboxylic acid cycle of Listeria.

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“…The complex media were L broth for routine subculture and peptone broth for specific growth tests (Guest, 1981), supplemented as required with ampicillin (100 pg ml-'), chloramphenicol (25 pg ml-'), kanamycin (15 pg ml-') and glucose (50 mM). The citrate-free minimal medium (Cole & Guest, 1980) was used with glucose (20 mM), sodium acetate (40 mM), sodium succinate (40 mM), sodium DL-lactate (40 mM) or sodium pyruvate (40 mM) as carbon sources, and supplements of thiamin hydrochloride (5 pg ml-l) and L-glutamate (2 mM) or L-proline (0.22 mM), as required.…”

Section: --mentioning

confidence: 99%

Two genetically-distinct and differentially-regulated aconitases (AcnA and AcnB) in Escherichia coli

1994

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Partial Replacement of Succinate Dehydrogenase Function by Phage- and Plasmid-specified Fumarate Reductase in Escherichia coli

Cited by 50 publications

References 11 publications

Aerobic inactivation of fumarate reductase from Escherichia coli by mutation of the [3Fe-4S]-quinone binding domain

Aerobic inactivation of fumarate reductase from Escherichia coli by mutation of the [3Fe-4S]-quinone binding domain

Functional γ-Aminobutyrate Shunt in Listeria monocytogenes: Role in Acid Tolerance and Succinate Biosynthesis

Two genetically-distinct and differentially-regulated aconitases (AcnA and AcnB) in Escherichia coli

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